5-Membered Annelated Heterocyclic Pyrimidines As Kinase Inhibitors

ABSTRACT

This invention comprises the novel compounds of formula (I)  
                 
 
wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , Q 1 , Q 2 , X 1 , X 2 , Y and Z have defined meanings, having cell cycle kinase inhibiting enzymatic activity, their preparation, compositions containing them and their use as a medicine.

FIELD OF THE INVENTION

The present invention relates to compounds and compositions containingsaid compounds acting as inhibitors of cell-cycle specific kinases, morein particular the cyclin dependent kinase CDK4 and/or the Aurora kinasesAURORA A and/or AURORA B. Moreover, the present invention providesprocesses for the preparation of the disclosed inhibitors, and methodsof using them, for instance as a medicine.

In normal cells the cell cycle is a tightly regulated and carefullybalanced process through which one cell divides into two. The fourphases, G1,S, G2 and M phase, reflect stages in cell cycle progressionwhere DNA synthesis and replication (S phase) and mitosis (M phase)occur in a temporally regulated fashion, separated by two gap phases (G1and G2). Cell cycle progression is maintained by an array of regulatorydecision points, governed in part by cyclin dependent kinases (Cdks),which determine whether it is appropriate for a cell to divide. Inaddition to the catalytic subunit (the Cdk itself), each Cdk complexcontains one of many activating subunits called cyclins because theirlevels fluctuate periodically throughout the cell cycle. Distinctcyclin-Cdk complexes power the cell through different phases of the cellcycle. In mammals, these complexes include the D-type cyclins whichactivate Cdk4 to execute critical regulatory events in G1; the E-typeand A-type cyclins, which activate Cdk2 to effect events in S phaseincluding DNA replication and centrosome duplication; and A-type cyclins(in a second role) and B-type cyclins, which activate Cdk1 to directstructural and regulatory events in mitosis. Inactivation of Cdk1 inlate mitosis contributes to reset the cell in G1.

An important role of Cdks is in the phosphorylation of theretinoblastoma (Rb) tumor suppression gene product whereafter E2F isreleased to facilitate DNA replication and progression through the cellcycle (McLaughlin et al., Drug Discovery Today. 8: 793-802 (2003)).

Cdk deregulation, either through direct or indirect mechanisms, is atypical feature in most cancer cells. Furthermore there exist a plethoraof biological mechanistic indications and convincing support bypreclinical studies, that Cdk inhibitors can synergise with variouschemotherapeutic agents in tumor cell killing (Fisher et al. ExpertOpin. Investig. Drugs. 12: 955-970 (2003)).

Also Aurora kinases play critical roles in cell division and chromosomesegregation. They are implicated in the centrosome cycle, spindleassembly, chromosome condensation, microtubule-kinetochore attachment,the spindle checkpoint and cytokinesis. Aurora kinases are regulatedthrough phosphorylation, the binding of specific partners andubiquitin-dependent proteolysis. The deregulation of Aurora kinasesimpairs spindle assembly, spindle checkpoint function and cell division,causing mis-segregation of individual chromosomes or polyploidizationaccompanied by centrosome amplification. Aurora kinases are frequentlyoverexpressed in cancers and the identification of Aurora A as acancer-susceptibility gene provides a strong link between mitotic errorsand carcinogenesis.

Thus pharmacological cell cycle specific kinase inhibition is anattractive strategy towards mechanism-based therapies in proliferativedisorders. Moreover, the combination of cell cycle specific kinasesinhibition with existing chemotherapy can have advantages effects.

BACKGROUND OF THE INVENTION

European patent application EP0961775 A2, published on 23 Apr. 1998,discloses purine L-nucleoside compounds and compositions that may beused in inflammation, infections, infestations, neoplasms, andautoimmune disease. More in particular these compounds are described asbeing modulators of Th1 and Th2.

European patent application EP 1147108 A1, published on 27 Jul. 2000,discloses substituted nitrogen heterocyclic derivatives havingimmunosuppressive, antimicrobial, cytostatic, anticancer, antimitoticand antineurogenerative effects. More in particular these compounds aredescribed as suppressors of spontaneous and mitogen activatedlymphocytes and as antiviral compounds. European patent applicationEP1244668 A1, published on 12 Jul. 2001, discloses purine derivativeswith an inhibitory effect on cyclin dependent kinases, viruses andproliferation of haematopoitic and cancer cells. More in particular thecompounds are described as inhibitors of the cyclin dependent kinasesthat associate with type B cyclin, f.e. cdk1 and related cdks (cdk2,cdk5, cdk7 and cdk9).

European patent application EP1507780, published on 4 Dec. 2003,discloses pyrazolo-pyrimidine aniline compounds, useful as kinaseinhibitors.

European application EP153976 A2, published on 4 Mar. 2004, describes2,6,9-trisubstituted 8-azapurines as kinase inhibitors.

European patent application EP1590341 A1, published on 5 Aug. 2004,discloses pyrimidine derivatives with an inhibitory activity oncyclin-dependent kinase 4.

European patent application EP1615926 A1, published on 4 Nov. 2004,discloses purin-6-yl amino acid derivatives as anti-cancer agents.

International application WO 03/63764, published on 9 Dec. 2004,discloses 6-substituted pyrazolo[3,4-d]pyrimidin-4-ones useful as cyclindependent kinase inhibitors

The present invention relates to compounds, which are distinguishablefrom the prior art in structure, pharmacological activity, potency andselectivity.

DESCRIPTION OF THE INVENTION

The present invention concerns a compound of formula (I)

a N-oxide, an addition salt, a quaternary amine and a stereochemic allyisomeric form thereof, wherein

X¹ and X² are each independently N or CH with the exception that X¹ andX² can not be both N;

Q¹ is CH₂, or N;

Q² is CH₂, N, or O;

n is an integer with value 0 or 1 and when n is 0 then a direct bond isintended;

t is an integer with value 0 or 1 and when t is 0 then a direct bond isintended;

-Y

Z- is —CH═N—, —C(═O)—NH—, —N═CH—, —C(═O)—CH₂—, or —NH—C(═O)—;

ring B represents phenyl, cyclopentyl, cyclohexyl, narbonyl or

-   -   L is a direct bond, —(CH₂)_(r)—NR⁷—(CH₂)_(s)—, —(CR⁸        ₂)_(r)—O—(CH₂)_(s)—, —C(═O)—, —(CH₂)_(r)—O—C(═O)—,        —(CH₂)_(r)—NR⁷—C(═O), —S(═O)₂—, —(CH₂)_(r)—NH—S(═O)₂—, or        —C₁₋₆alkyl-; wherein        -   each —(CH₂)_(r)— moiety is linked to ring A;        -   each s is an integer with value 0 or 1 and when s is 0 then            a direct bond is intended; each r is an integer with value            0, 1, 2 or 3 and when r is 0 then a direct bond is intended;        -   each R⁷ is hydrogen, C₁₋₆alkyl or C₁₋₄alkyloxycarbonyl;        -   each R⁸ is independently hydrogen, hydroxy or C₁₋₆alkyl; or        -   two R⁸ together can form a bivalent radical of formula            —CH₂—CH₂—;    -   R¹, R² and R⁵ are each independently hydrogen, hydroxy or        C₁₋₆alkyl;    -   R³ is hydrogen, hydroxy, C₁₋₆alkyl, hydroxyC₁₋₆alkyl,        hydroxycyclopropylC₁₋₆alkyl, hydroxycyclopropylcarbonyl,        hydroxyC₁₋₆alkylcarbonyl, hydroxyC₁₋₆alkyloxy, C₁₋₆alkyloxy,        hydroxyC₁₋₆alkyloxyC₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkylcarbonyl,        C₁₋₄alkyloxycarbonyl, C₁₋₆alkyloxyC₁₋₆alkyl,        C₁₋₆alkyloxyC₁₋₆alkylcarbonyl, C₁₋₆alkyloxyC₁₋₆alkyloxy,        C₁₋₆alkyloxyC₁₋₆alkyloxyC₁₋₆alkyl, pyridinyl, —NR⁹R¹⁰, or        —S(═O)₂—NR⁹R¹⁰;        -   wherein each R⁹ and R¹⁰ independently represent hydrogen,            C₁₋₆alkyl, C₁₋₄alkyloxycarbonyl, hydroxyC₁₋₆alkyl,            hydroxycyclopropylC₁₋₆alkyl or C₁₋₆alkyloxyC₁₋₆alkyl;    -   R⁴ is hydrogen or halo;    -   R⁶ is hydrogen, C₁₋₆alkyl or C₁₋₄alkyloxycarbonyl.

The compounds of formula (I) may also exist in their tautomeric forms.Such forms although not explicitly indicated in the above formula areintended to be included within the scope of the present invention.

A number of terms used in the foregoing definitions and hereinafter areexplained hereunder. These terms are sometimes used as such or incomposite terms.

As used in the foregoing definitions and hereinafter, halo is generic tofluoro, chloro, bromo and iodo; C₁₋₄alkyl defines straight and branchedchain saturated hydrocarbon radicals having from 1 to 4 carbon atomssuch as, e.g. methyl, ethyl, propyl, butyl, 1-methylethyl,2-methylpropyl and the like; C₁₋₆alkyl includes C₁₋₄alkyl and the higherhomologues thereof having 5 to 6 carbon atoms such as, for example,pentyl, 2-methyl-butyl, hexyl, 2-methylpentyl and the like.

The term “addition salt” comprises the salts which the compounds offormula (I) are able to form with organic or inorganic bases such asamines, alkali metal bases and earth alkaline metal bases, or quaternaryammonium bases, or with organic or inorganic acids, such as mineralacids, sulfonic acids, carboxylic acids or phosphorus containing acids.

The term “addition salt” further comprises pharmaceutically acceptablesalts, metal complexes and solvates and the salts thereof, that thecompounds of formula (I) are able to form.

The term “pharmaceutically acceptable salts” means pharmaceuticallyacceptable acid or base addition salts. The pharmaceutically acceptableacid or base addition salts as mentioned hereinabove are meant tocomprise the therapeutically active non-toxic acid and non-toxic baseaddition salt forms which the compounds of formula (I) are able to form.The compounds of formula (I) which have basic properties can beconverted in their pharmaceutically acceptable acid addition salts bytreating said base form with an appropriate acid. Appropriate acidscomprise, for example, inorganic acids such as hydrohalic acids, e.g.hydrochloric or hydrobromic acid; sulfuric; nitric; phosphoric and thelike acids; or organic acids such as, for example, acetic, propanoic,hydroxyacetic, lactic, pyruvic, oxalic, malonic, succinic (i.e.butanedioic acid), maleic, fumaric, malic, tartaric, citric,methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluenesulfonic,cyclamic, salicylic, p-aminosalicylic, pamoic and the like acids.

The compounds of formula (I) which have acidic properties may beconverted in their pharmaceutically acceptable base addition salts bytreating said acid form with a suitable organic or inorganic base.Appropriate base salt forms comprise, for example, the ammonium salts,the alkali and earth alkaline metal salts, e.g. the lithium, sodium,potassium, magnesium, calcium salts and the like, salts with organicbases, e.g. the benzathine, N-methyl-D-glucamine, hydrabamine salts, andsalts with amino acids such as, for example, arginine, lysine and thelike.

The terms acid or base addition salt also comprise the hydrates and thesolvent addition forms which the compounds of formula (I) are able toform. Examples of such forms are e.g. hydrates, alcoholates and thelike.

The term “metal complexes” means a complex formed between a compound offormula (I) and one or more organic or inorganic metal salt or salts.Examples of said organic or inorganic salts comprise the halogenides,nitrates, sulfates, phosphates, acetates, trifluoroacetates,trichloroacetates, propionates, tartrates, sulfonates, e.g.methylsulfonates, 4-methylphenylsulfonates, salicylates, benzoates andthe like of the metals of the second main group of the periodicalsystem, e.g. the magnesium or calcium salts, of the third or fourth maingroup, e.g. aluminium, tin, lead as well as the first to the eighthtransition groups of the periodical system such as, for example,chromium, manganese, iron, cobalt, nickel, copper, zinc and the like.

The term “stereochemically isomeric forms of compounds of formula (I)”,as used hereinbefore, defines all possible compounds made up of the sameatoms bonded by the same sequence of bonds but having differentthree-dimensional structures which are not interchangeable, which thecompounds of formula (I) may possess. Unless otherwise mentioned orindicated, the chemical designation of a compound encompasses themixture of all possible stereochemically isomeric forms which saidcompound may possess. Said mixture may contain all diastereomers and/orenantiomers of the basic molecular structure of said compound. Allstereochemically isomeric forms of the compounds of formula (I) both inpure form or in admixture with each other are intended to be embracedwithin the scope of the present invention.

The N-oxide forms of the compounds of formula (I) are meant to comprisethose compounds of formula (I) wherein one or several nitrogen atoms areoxidized to the so-called N-oxide, particularly those N-oxides whereinone or more of the piperidine-, piperazine or pyridazinyl-nitrogens areN-oxidized.

Whenever used hereinafter, the term “compounds of formula (I)” is meantto include also the N-oxide forms, the pharmaceutically acceptable acidor base addition salts and all stereoisomeric forms.

As used herein before, the term (═O) forms a carbonyl moiety whenattached to a carbon atom, a sulfoxide moiety when attached to a sulfuratom and a sulfonyl moiety when two of said terms are attached to asulfur atom.

Lines drawn into ring systems from substituents indicate that the bondmay be attached to any of the suitable ring atoms of the ring system.

A first group of interesting compounds consists of those compounds offormula (I) wherein one or more of the following restrictions apply:

-   -   a) n is 0;    -   b) t is 0;    -   c) L is a direct bond, —(CH₂)_(r)—NR⁷—(CH₂)_(s)—, —C(═O)—,        —(CH₂)_(r)—NR⁷—C(═O)—, —S(═O)₂— or —C₁₋₆alkyl-;    -   d) s is 1;    -   e) r is 0 or 2;    -   f) each R⁷ is hydrogen or C₁₋₄alkyloxycarbonyl;    -   g) R¹, R² and R⁵ are each independently hydrogen;    -   h) R³ is hydrogen, hydroxy, C₁₋₆alkyl, hydroxyC₁₋₆alkyl,        hydroxycyclopropylcarbonyl, hydroxyC₁₋₆alkylcarbonyl,        C₁₋₆alkylcarbonyl, C₁₋₄alkyloxycarbonyl, C₁₋₆alkyloxyC₁₋₆alkyl,        C₁₋₆alkyloxyC₁₋₆alkylcarbonyl, pyridinyl, —NR⁹R¹⁰, or        —S(═O)₂—NR⁹R¹⁰;    -   i) each R⁹ and R¹⁰ independently represent hydrogen,        C₁₋₄alkyloxycarbonyl or C₁₋₆alkyloxyC₁₋₆alkyl; and    -   j) R⁶ is hydrogen or C₁₋₄alkyloxycarbonyl.

A second group of interesting compounds consists of those compounds offormula (I) wherein one or more of the following restrictions apply:

-   -   a) X¹ and X² are each CH;    -   b) Q² is N, or O;    -   c) n is 0;    -   d) t is 0;    -   e) -Y        Z- is —N═CH—;    -   f) ring B represents cyclohexyl or narbonyl;    -   g) L is a direct bond, —(CH₂)_(r)—NR⁷—(CH₂)_(s)—, —C(═O)— or        —C₁₋₆alkyl-;    -   h) s is 1;    -   i) r is 0 or 2;    -   j) each R⁷ is hydrogen;    -   k) R¹, R² and R⁵ are each independently hydrogen;    -   l) R³ is hydrogen, C₁₋₆alkyl or hydroxyC₁₋₆alkyl;    -   m) R⁴ is hydrogen; and    -   n) R⁶ is hydrogen.

A third group of interesting compounds consists of those compounds offormula (I) wherein one or more of the following restrictions apply:

-   -   a) X¹ and X² are each CH;    -   b) Q¹ is N;    -   c) Q² is N;    -   d) n is 0;    -   e) t is 0;    -   f) -Y        Z- is —N═CH—;    -   g) L is a direct bond or methyl;    -   h) R¹, R² and R⁵ are each independently hydrogen;    -   k) R³ is C₁₋₆alkyl;    -   l) each R⁹ and R¹⁰ independently represent hydrogen,        C₁₋₄alkyloxycarbonyl, C₁₋₆alkyloxyC₁₋₆alkyl;    -   m) R⁴ is hydrogen; and    -   n) R⁶ is hydrogen.

A fourth group of interesting compounds consists of those compounds offormula (I) and those compounds of the above mentioned groups wherein -Y

Z- is —N═CH—.

A fifth group of interesting compounds consists of those compounds offormula (I) and those compounds of the above mentioned groups whereinring B represents cyclohexyl or narbonyl.

A sixth group of interesting compounds consists of those compounds offormula (I) and those compounds of the above mentioned groups wherein Q¹is N and L is a direct bond, —C(═O)—, or methyl.

A seventh group of interesting compounds consists of those compounds offormula (I) and those compounds of the above mentioned groups wherein Q¹is CH₂ and L is —(CH₂)_(r)—NR⁷—CH₂—.

A ninth group of interesting compounds consists of those compounds offormula (I) and those compounds of the above mentioned groups whereinR³, R⁶, R⁷, R⁹, R¹⁰, are other than C₁₋₄alkyloxycarbonyl.

A tenth group of interesting compounds consists of those compounds offormula (I) and those compounds of the above mentioned groups wherein X¹and X² are each CH.

A group of preferred compounds consists of those compounds of formula(I) wherein n is 0; t is 0; L is a direct bond,—(CH₂)_(r)—NR⁷—(CH₂)_(s)—, —C(═O)—, —(CH₂)_(r)—NR⁷—C(═O)—, —S(═O)₂— or—C₁₋₆alkyl-; s is 1; r is 0 or 2; each R⁷ is hydrogen orC₁₋₄alkyloxycarbonyl; R¹, R² and R⁵ are each independently hydrogen; R³is hydrogen, hydroxy, C₁₋₆alkyl, hydroxyC₁₋₆alkyl,hydroxycyclopropylcarbonyl, hydroxyC₁₋₆alkylcarbonyl, C₁₋₆alkylcarbonyl,C₁₋₄alkyloxycarbonyl, C₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkyloxyC₁₋₆alkylcarbonyl, pyridinyl, —NR⁹R¹⁰, or —S(═O)₂—NR⁹R¹⁰;each R⁹ and R¹⁰ independently represent hydrogen, C₁₋₄alkyloxycarbonylor C₁₋₆alkyloxyC₁₋₆alkyl; and R⁶ is hydrogen or C₁₋₄alkyloxycarbonyl.

A group of more preferred compounds consists of those compounds offormula (I) wherein X¹ and X² are each CH; Q² is N, or O; n is 0; t is0; -Y

Z- is —N═CH—; ring B represents cyclohexyl or narbonyl; L is a directbond, —(CH₂)_(r)—NR⁷—(CH₂)_(s)—, —C(═O)— or —C₁₋₆alkyl-; s is 1; r is 0or 2; each R⁷ is hydrogen; R¹, R² and R⁵ are each independentlyhydrogen; R³ is hydrogen, C₁₋₆alkyl or hydroxyC₁₋₆alkyl; R⁴ is hydrogen;and R⁶ is hydrogen.

Another group of more preferred compounds consists of those compounds offormula (I) wherein X¹ and X² are each CH; Q² is N, or O; n is 0; t is0; -Y

Z- is —N═CH—; ring B represents cyclohexyl or narbonyl; L is a directbond, —NH—CH₂—, —C(═O)— or methyl; R¹, R² and R⁵ are each independentlyhydrogen; R⁴ is hydrogen; and R⁶ is hydrogen.

The most preferred compounds are compound No 1, compound No 2, compoundNo 15, compound No 3, compound No 60, compound No 37, compound No 4 andcompound No 23.

Compound No 1

Compound No 2

Compound No 15

Compound No 3

Compound No 60

Compound No 37

Compound No 4

Compound No 23

The compounds of formula (I), their pharmaceutically acceptable saltsand N-oxides and stereochemically isomeric forms thereof may be preparedin conventional manner. The starting materials and some of theintermediates are known compounds and are commercially available or maybe prepared according to conventional reaction procedures as generallyknown in the art.

A number of such preparation methods will be described hereinafter inmore detail.

Compounds of formula (I) can be prepared by reacting an intermediate offormula (II) with an intermediate of formula (III) in the presence of asuitable solvent, such as for example (CH₃)₂N—C(═O)H, dimethylsulfoxide,CH₃—O—CH₂—CH₂—OH, an alcohol, e.g. 2-propanol and the like, optionallyin the presence of a suitable base, such as for exampleN,N-diisopropylethanamine, NaH or 2,6-dimethylpyridine.

Compounds of formula (I) can also be prepared by reacting anintermediate of formula (X), wherein W¹ is a suitable leaving group suchas halogen, for example chloro and the like, with an intermediate offormula (II) in the presence of a suitable solvent, such as for example(CH₃)₂N—C(═O)H, dimethylsulfoxide, CH₃—O—CH₂—CH₂—OH or an alcohol, e.g.2-propanol and the like, optionally in the presence of a suitable base,such as for example N,N-diisopropylethanamine, NaH or2,6-dimethylpyridine.

Compounds of formula (I) wherein L is —(CH₂)_(r)—NH—C(═O)—, hereinreferred to as compounds of formula (I-a), can also be prepared byreacting an intermediate of formula (XIV) with an intermediate offormula (XVI-a) in the presence of a suitable solvent, such as forexample (CH₃)₂N—C(═O)H, dimethylsulfoxide, CH₃—O—CH₂—CH₂—OH or analcohol, e.g. 2-propanol and the like, optionally in the presence of asuitable base, such as for example N,N-diisopropylethanamine, NaH or2,6-dimethylpyridine.

Compounds of formula (I) wherein L is —(CH₂)_(r)—NH—(CH₂)—, hereinreferred to as compounds of formula (I-b) can also be prepared byreacting intermediates of formula (XIV) with intermediates of formula(XVI-b) in the presence of sodium triacetoxyborohydride or sodiumcyanoborohydride, in the presence of a suitable acid, such as aceticacid and in a suitable solvent, such as for example methanol ortetrahydrofuran.

Compounds of formula (I), wherein -Y

Z- is —CH═N—, herein referred to as compounds of formula (I-c) can beprepared by cyclizing an intermediate of formula (XXXIII) in thepresence of a salt, such as for example Gold's reagent, or an acid, suchas for example formic acid, in a suitable solvent, such as for exampledioxane or thriethoxymethane

In the above reactions, the obtained compound of formula (I) can beisolated, and, if necessary, purified according to methodologiesgenerally known in the art such as, for example, extraction,crystallization, distillation, trituration and chromatography. In casethe compound of formula (I) crystallizes out, it can be isolated byfiltration. Otherwise, crystallization can be caused by the addition ofan appropriate solvent, such as for example water; acetonitrile; analcohol, such as for example methanol; and combinations of saidsolvents. Alternatively, the reaction mixture can also be evaporated todryness, followed by purification of the residue by chromatography (e.g.reverse phase HPLC, flash chromatography and the like). The reactionmixture can also be purified by chromatography without previouslyevaporating the solvent. The compound of formula (I) can also beisolated by evaporation of the solvent followed by recrystallisation inan appropriate solvent, such as for example water; acetonitrile; analcohol, such as for example methanol; and combinations of saidsolvents. The person skilled in the art will recognise which methodshould be used, which solvent is the most appropriate to use or itbelongs to routine experimentation to find the most suitable isolationmethod.

In this and the following preparations, the reaction products may beisolated from the reaction medium and, if necessary, further purifiedaccording to methodologies generally known in the art such as, forexample, extraction, crystallization, distillation, trituration andchromatography.

The compounds of formula (I) may also be converted into each other viaart-known reactions or functional group transformations. A number ofsuch transformations are already described hereinabove. Other examplesare hydrolysis of carboxylic esters to the corresponding carboxylic acidor alcohol; hydrolysis of amides to the corresponding carboxylic acidsor amines; hydrolysis of nitriles to the corresponding amides; aminogroups on imidazole or phenyl may be replaced by a hydrogen by art-knowndiazotation reactions and subsequent replacement of the diazo-group byhydrogen; alcohols may be converted into esters and ethers; primaryamines may be converted into secondary or tertiary amines; double bondsmay be hydrogenated to the corresponding single bond; an iodo radical ona phenyl group may be converted in to an ester group by carbon monoxideinsertion in the presence of a suitable palladium catalyst.

The compounds of formula (I) may be converted to the correspondingN-oxide forms following art-known procedures for converting a trivalentnitrogen into its N-oxide form. Said N-oxidation reaction may generallybe carried out by reacting the starting material of formula (I) with anappropriate organic or inorganic peroxide. Appropriate inorganicperoxides comprise, for example, hydrogen peroxide, alkali metal orearth alkaline metal peroxides, e.g. sodium peroxide, potassiumperoxide; appropriate organic peroxides may comprise peroxy acids suchas, for example, benzenecarboper-oxoic acid or halo substitutedbenzenecarboperoxoic acid, e.g. 3-chlorobenzenecarbo-peroxoic acid,peroxoalkanoic acids, e.g. peroxoacetic acid, alkylhydroperoxides, e.g.t.butyl hydro-peroxide. Suitable solvents are, for example, water, loweralcohols, e.g. ethanol and the like, hydrocarbons, e.g. toluene,ketones, e.g. 2-butanone, halogenated hydrocarbons, e.g.dichloromethane, and mixtures of such solvents.

Some of the compounds of formula (I) and some of the intermediates inthe present invention may consist of a mixture of stereochemicallyisomeric forms. Pure stereochemically isomeric forms of said compoundsand said intermediates can be obtained by the application of art-knownprocedures. For example, diastereoisomers can be separated by physicalmethods such as selective crystallization or chromatographic techniques,e.g. counter current distribution, liquid chromatography and the likemethods. Enantiomers can be obtained from racemic mixtures by firstconverting said racemic mixtures with suitable resolving agents such as,for example, chiral acids, to mixtures of diastereomeric salts orcompounds; then physically separating said mixtures of diastereomericsalts or compounds by, for example, selective crystallization orchromatographic techniques, e.g. liquid chromatography and the likemethods; and finally converting said separated diastereomeric salts orcompounds into the corresponding enantiomers. Pure stereochemicallyisomeric forms may also be obtained from the pure stereochemicallyisomeric forms of the appropriate intermediates and starting materials,provided that the intervening reactions occur stereospecifically.

An alternative manner of separating the enantiomeric forms of thecompounds of formula (I) and intermediates involves liquidchromatography, in particular liquid chromatography using a chiralstationary phase.

Intermediates of formula (XVI-a) can be prepared by converting of anintermediate of formula (XVII) in the presence of a suitable acid suchas trifluoroacetic acid in a suitable solvent such as CH₂C₂

Intermediates of formula (XVII) can be prepared by reacting anintermediate of formula (XVIII) with an intermediate of formula (III) inthe presence of a suitable solvent, such as for example (CH₃)₂N—C(═O)H,dimethylsulfoxide, CH₃—O—CH₂—CH₂—OH or an alcohol, e.g. 2-propanol andthe like, optionally in the presence of a suitable base, such as forexample N,N-diisopropylethanamine, Cesium carbonate, NaH or2,6-dimethylpyridine.

Intermediates of formula (XXXIII) can be prepared by reducing anintermediate of formula (XXXIV) with a suitable reducing agent, such asfor example H₂, in the presence of a suitable catalyst, such as forexample platina on charcoal or palladium on charcoal, optionally in thepresence of a suitable catalyst poison, such as for example a thiophenesolution, optionally in the presence of NH₂—NH₂, in the presence of asuitable solvent, such as for example N,N-dimethylacetamide,tetrahydrofuran, N,N-dimethylformamide or a suitable alcohol, such asfor example methanol, ethanol and the like, and optionally in thepresence of a suitable base, such as for example N,N-diethylethanamine.

Intermediates of formula (XXXIV) can also be prepared by reacting anintermediate of formula (XXXV) wherein W¹ represents a suitable leavinggroup such as halogen, for example chloro and the like, with anintermediate of formula (II) in the presence of a suitable base, such asfor example N,N-diisopropylethanamine or N,N-diethylethanamine, andoptionally in the presence of a suitable solvent, such as for exampleN,N-dimethylacetamide, N,N-dimethylformamide or 1,4-dioxane.

Intermediates of formula (III) can be prepared by reacting anintermediate of formula (IV) with a suitable oxidizing agent, such asfor example KMnO₄ in the presence of a suitable solvent, such as forexample water, and a suitable acid, such as for example acetic acid. Analternative suitable oxidizing agent is meta-chloroperbenzoic acidoptionally in the presence of MgSO₄, in a suitable solvent, such as forexample CH₂Cl₂ and optionally an alcohol, e.g. methanol and the like,optionally in the presence of morpholinomethyl PS and PS-ammoniumbicarbonate scavenger.

Intermediates of formula (IV), wherein -Y

Z- is —N═CH—, herein referred to as intermediates of formula (IV-a) canbe prepared by cyclizing an intermediate of formula (V) with a suitableacid, such as for example HCl and the like, in the presence of asuitable solvent, such as for example 2-propanol.

Intermediates of formula (IV), wherein -Y

Z- is —NH—C(═O)—, herein referred to as intermediates of formula (IV-b)can be prepared by cyclizing an intermediate of formula (VII) with asuitable acid, such as for example HCl and the like, in the presence ofa suitable solvent, such as for example 2-propanol.

Intermediates of formula (IV), wherein -Y

Z- is —C(═O)—CH₂—, herein referred to as intermediates of formula (IV-c)can be prepared by cyclizing an intermediate of formula (XIX) in thepresence of N,N′-methanetetraylbis-2-propanamine and a suitable solvent,such as for example CH₂Cl₂.

Intermediates of formula (IV), wherein -Y

Z- is —C(═O)—CH₂—, herein referred to as intermediates of formula (IV-d)can be prepared by cyclizing an intermediate of formula (XXV) in thepresence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and a suitablesolvent, such as for example CH₂Cl₂, in the presence of a suitable base,such as for example N,N-diisopropylethanamine.

Intermediates of formula (V) can be prepared by reacting an intermediateof formula (VI) with a suitable agent, such as for example MnO₂ in thepresence of a suitable solvent, such as for example CH₂Cl₂.

Intermediates of formula (VI) can be prepared by reacting anintermediate of formula (VII) with a suitable agent, such as for exampleLiAlH₄ in the presence of a suitable solvent, such as for exampletetrahydrofuran.

Intermediates of formula (VII) can be prepared by reacting anintermediate of formula (VIII), wherein W² represents a suitable leavinggroup such as halogen, for example. chloro and the like, with anintermediate of formula (IX) in the presence of a suitable solvent, suchas for example tetrahydrofuran, in the presence of a suitable base, suchas for example N,N-diethylethanamine.

Intermediates of formula (XIX) can be prepared by reducing anintermediate of formula (XX) with a suitable reducing agent, such as forexample H₂, in the presence of a suitable catalyst, such as for exampleplatina on charcoal or palladium on charcoal, optionally in the presenceof a suitable catalyst poison, such as for example a thiophene solution,optionally in the presence of NH₂—NH₂, in the presence of a suitablesolvent, such as for example N,N-dimethylacetamide, tetrahydrofuran,N,N-dimethylformamide or a suitable alcohol, such as for examplemethanol, ethanol and the like, and optionally in the presence of asuitable base, such as for example N,N-diethylethanamine.

Intermediates of formula (XX) can be prepared by reacting anintermediate of formula (XXI), wherein W² represents a suitable leavinggroup such as halogen, for example, chloro and the like, with anintermediate of formula (XXII) in the presence of a suitable base, suchas for example N,N-diisopropylethanamine or N,N-diethylethanamine, andoptionally in the presence of a suitable solvent, such as for exampleN,N-dimethylacetamide, N,N-dimethylformamide, acetonitrile or1,4-dioxane.

Intermediates of formula (XXI) can be prepared by reacting anintermediate of formula (XXIII) with POCl₃.

Intermediates of formula (XXIII) can be prepared by reacting anintermediate of formula (XXIV), wherein W¹ represents a suitable leavinggroup such as halogen, for example, chloro and the like, in the presenceof NaS—CH₃ in water, in a suitable solvent, such as for example ethanol.

Intermediates of formula (XXV) can be prepared by reacting anintermediate of formula (XXVI) with a suitable acid, such as for exampleHCl in the presence of a suitable solvent, such as for example dioxane.

Intermediates of formula (XXVI) can be prepared by reacting anintermediate of formula (XXVII) wherein W² represents a suitable leavinggroup such as halogen, for example chloro and the like, with anintermediate of formula (XXVIII) in the presence of a suitable solvent,such as for example cyclohexylamine, in the presence of a suitable base,such as for example N,N-diethylethanamine.

Intermediates of formula (XXVII), wherein halo represents for examplechloro and the like, can be prepared by reacting an intermediate offormula (XXIX) with POCl₃.

Intermediates of formula (XXIX) can be prepared by reacting anintermediate of formula (XXX) with iodomethane in the presence of asuitable solvent such as for example a mixture of2,3,4,6,7,8,9,10-octahydro-pyrimido[1,2-a]azepine andN,N-dimethylformamide.

Intermediates of formula (XXX) can be prepared by reacting anintermediate of formula (XXXI) with thiourea in the presence of sodiumethoxylate in a suitable solvent such as ethanol.

Intermediates of formula (XXXI) can be prepared by reacting anintermediate of formula (XXXII) with H(C═O)—O—CH₂—CH₃ in the presence ofNaH in a suitable solvent such as a mixture of ethanol andtetrahydrofuran.

Intermediates of formula (II) wherein R⁶ is C₁₋₄alkyloxycarbonyl, suchas tertiary butoxycarbonyl, herein referred to as intermediates offormula (II-a), can be prepared by reacting intermediates of formula(II-b) with di-tert-butyl-dicarbonate in the presence of a suitablesolvent, such as for example tetrahydrofuran, and optionally in thepresence of a suitable base, such as for example N,N-diethylethanamine.

Intermediates of formula (II) wherein R⁶ represents hydrogen, saidintermediates being represented by formula (II-b), can be prepared byreacting an intermediate of formula (XI) with a suitable reducing agent,such as for example H₂, in the presence of a suitable catalyst, such asfor example platina on charcoal or palladium on charcoal, optionally asuitable catalyst poison, such as for example a thiophene solution, asuitable solvent, such as for example N,N-dimethylacetamide,tetrahydrofuran, N,N-dimethylformamide or a suitable alcohol, such asfor example methanol, and optionally in the presence of a suitable base,such as for example N,N-diethylethanamine.

Intermediates of formula (XI) can be prepared by reacting anintermediate of formula (XII), wherein W³ represents a suitable leavinggroup such as halogen, for example, chloro and the like, with anintermediate of formula (XIII) in the presence of a suitable solvent,such as for example N,N-dimethylacetamide, N,N-dimethylformamide,CH₂Cl₂, 1,4-dioxane, or a mixture of CH₂Cl₂ and pyridine and optionallyin the presence of a suitable base, such as for exampleN,N-diisopropylethanamine.

Intermediates of formula (XI) wherein L is —(CH₂)_(r)—NR⁷—(CH₂)— and R⁷is C₁₋₄alkyloxycarbonyl, such as f.e. tertiary butoxycarbonyl, hereinreferred to as intermediates of formula (XI-a), can be prepared byreacting an intermediate of formula (XI) wherein L is—(CH₂)_(r)—NR⁷—(CH₂)— and R⁷ is hydrogen, herein referred to asintermediates of formula (XI-b), with di-tert-butyl-dicarbonate in thepresence of a suitable solvent, such as for example tetrahydrofuran, andoptionally in the presence of a suitable base, such as for exampleN,N-diethylethanamine.

Intermediates of formula (XI-b) can be prepared by reactingintermediates of formula (XIV) with intermediates of formula (XV) in thepresence of sodium triacetoxyborohydride or sodium cyanoborohydride, inthe presence of a suitable acid, such as acetic acid and in a suitablesolvent, such as for example methanol or tetrahydrofuran.

The compounds of formula (I), the pharmaceutically acceptable acidaddition salts and stereoisomeric forms thereof have valuablepharmacological properties in that they are selective cell cyclespecific kinase inhibitors. Specific inhibitory compounds are superiortherapeutic agents since they are characterized by a greater efficacyand lower toxicity by virtue of their specificity.

The term “cell cycle specific kinase inhibitor(s)” or “inhibitor(s) ofcell-cycle specific kinases” is used herein to describe an agent thatinhibits at least the activity of CDK4, AURORA A and/or AURORA B in theassays described in C (pharmacological examples).

The term “CDK4” is used herein to mean a protein obtained as a result ofexpression of a cdk4 gene. Within the meaning of this term, CDK4encompass all proteins encoded by a cdk4 gene, mutants thereof, andalternative slice proteins thereof. Additionally, as used herein, theterm “CDK4” includes CDK4 analogues, homologues and analogues of otheranimals.

The term “AURORA A and/or AURORA B” is used herein to mean a proteinobtained as a result of expression of an aurora gene. Within the meaningof this term, AURORA A and/or AURORA B encompass all proteins encoded byan aurora gene, mutants thereof, and alternative slice proteins thereof.Additionally, as used herein, the term “AURORA A and/or AURORA B”includes AURORA A and/or AURORA B analogues, homologues and analogues ofother animals.

The term “cell cycle specific kinase”, includes, but is not limited to,cyclin dependent kinases and/or aurora kinases.

The term “cyclin dependent kinases”, includes but is not limited toCDK4. Within the meaning of this term CDK1, CDK2, CDK3, CDK5, CDK6,CDK7, CDK8 and CDK9 may be encompassed.

Hence, the present invention discloses the compounds of formula (I) foruse as a medicine.

Furthermore, the invention also concerns the use of a compound for themanufacture of a medicament for the treatment of a disorder mediatedthrough cell cycle specific kinases.

In particular, the invention concerns the use of a compound for themanufacture of a medicament for the treatment of a disorder mediatedthrough CDK4, AURORA A and/or AURORA B.

Even more in particular, the invention concerns the use of a compoundfor the manufacture of a medicament for the treatment of a proliferativedisorder or a differentiative disorder.

The term “proliferative disorder” is used herein in a broad sense toinclude any disorder that requires control of the cell cycle, forexample cancer; cardiovascular disorders such as f.e. restenosis andcardiomyopathy; auto-immune disorders such as f.e. glomerulonephritis,rheumatoid arthritis, lupus, type I diabetis, and multiple sclerosis;dermatological disorders such as f.e. psoriasis, anti-inflammatorydisorders and anti-viral disorders. In these disorders, the compounds offormula (I) may induce apoptosis or maintain stasis within the desiredcells as required.

The compounds of the invention may inhibit any of the steps or stages inthe cell cycle, such as for example:

-   -   formation of the nuclear envelope,    -   exit from the quiescent phase of the cell cycle (G0),    -   G1 entry,    -   G1 progression,    -   chromosome decondensation,    -   nuclear envelope breakdown,    -   START,    -   initiation of DNA replication,    -   progression of DNA replication,    -   termination of DNA replication,    -   centrosome duplication,    -   G2 entry,    -   G2 progression,    -   activation of mitotic or meiotic function,    -   chromosome condensation,    -   centrosome separation,    -   microtubule nucleation,    -   spindle formation and/or function,    -   interaction with microtubule motor proteins,    -   chromatid separation and segregation,    -   inactivation of mitotic function,    -   formation of the contractile ring, and/or    -   cytokinesis functions.

The compounds of the invention may in particular inhibit replication ofRNA and DNA viruses that are dependent upon events associated with hostcell proliferation and differentiation.

With the term “differentiative disorder” is meant any disorder thatresults from the de-differentiation of tissue which may (optionally) beaccompanied by re-entry in mitosis. Such degenerative disorders includeneurodegenerative diseases of the nervous system, such as f.e.,Alzheimer's disease, Parkinson's disease, Huntington's disease, oramyotrophic lateral sclerosis as well as spinocerebellar degenerations.Other differentiative disorders include, for example, disordersassociated with connective tissue such as may occur due tode-differentiation of chondrocytes or osteocytes, cardiovasculardisorders which involve the de-differentiation of endothelial tissue andsmooth muscle cells, gastric ulcers characterised by degenerativechanges in glandular cells, and renal conditions marked by failure todifferentiate.

The term “treating” or “treatment” as used herein covers any treatmentof a disease and/or condition in an animal, particularly a human, andincludes: (i) preventing a disease and/or condition from occurring in asubject which may be predisposed to the disease and/or condition but hasnot yet been diagnosed as having it; (ii) inhibiting the disease and/orcondition, i.e., arresting its development; (iii) relieving the diseaseand/or condition, i.e., causing regression of the disease and/orcondition.

This invention also provides a method for treating a disorder mediatedthrough a cell cycle specific kinase by administering an effectiveamount of a compound of the present invention, to a subject, e.g. amammal (and more particularly a human) in need of such treatment.

In particular, this invention provides a method for treating a disordermediated through CDK4, AURORA A and/or AURORA B, by administering aneffective amount of a compound of the present invention, to a subject,e.g. a mammal (and more particularly a human) in need of such treatment.

Even more in particular, this invention provides a method for treating aproliferative and/or a differentiative disorder, by administering aneffective amount of a compound of the present invention, to a subject,e.g. a mammal (and more particularly a human) in need of such treatment.

Thus, this invention also provides a method for inhibiting tumour growthby administering an effective amount of a compound of the presentinvention, to a subject, e.g. a mammal (and more particularly a human)in need of such treatment.

Examples of tumours which may be inhibited, but are not limited to, lungcancer (e.g. adenocarcinoma and including non-small cell lung cancer),pancreatic cancers (e.g. pancreatic carcinoma such as, for exampleexocrine pancreatic carcinoma), colon cancers (e.g. colorectalcarcinomas, such as, for example, colon adenocarcinoma and colonadenoma), oesophageal cancer, oral squamous carcinoma, tongue carcinoma,gastric carcinoma, nasopharyngeal cancer, hematopoietic tumours oflymphoid lineage (e.g. acute lymphocytic leukemia, B-cell lymphoma,Burkitt's lymphoma), myeloid leukemias (for example, acute myelogenousleukemia (AML)), thyroid follicular cancer, myelodysplastic syndrome(MDS), tumours of mesenchymal origin (e.g. fibrosarcomas andrhabdomyosarcomas), melanomas, teratocarcinomas, neuroblastomas, braintumors, gliomas, benign tumour of the skin (e.g. keratoacanthomas),breast carcinoma (e.g. advanced breast cancer), kidney carcinoma, ovarycarcinoma, cervical carcinoma, endometrial carcinoma, bladder carcinoma,prostate cancer including the advanced disease, testicular cancers,osteosarcoma, head and neck cancer and epidermal carcinoma.

In view of their useful pharmacological properties, the subjectcompounds may be formulated into various pharmaceutical forms foradministration purposes.

To prepare the pharmaceutical compositions of this invention, aneffective amount of a particular compound, in base or acid addition saltform, as the active ingredient is combined in intimate admixture with apharmaceutically acceptable carrier, which carrier may take a widevariety of forms depending on the form of preparation desired foradministration. These pharmaceutical compositions are desirably inunitary dosage form suitable, preferably, for administration orally,rectally, percutaneously, or by parenteral injection. For example, inpreparing the compositions in oral dosage form, any of the usualpharmaceutical media may be employed, such as, for example, water,glycols, oils, alcohols and the like in the case of oral liquidpreparations such as suspensions, syrups, elixirs and solutions; orsolid carriers such as starches, sugars, kaolin, lubricants, binders,disintegrating agents and the like in the case of powders, pills,capsules and tablets.

Because of their ease in administration, tablets and capsules representthe most advantageous oral dosage unit form, in which case solidpharmaceutical carriers are obviously employed. For parenteralcompositions, the carrier will usually comprise sterile water, at leastin large part, though other ingredients, to aid solubility for example,may be included. Injectable solutions, for example, may be prepared inwhich the carrier comprises saline solution, glucose solution or amixture of saline and glucose solution. Injectable suspensions may alsobe prepared in which case appropriate liquid carriers, suspending agentsand the like may be employed. In the compositions suitable forpercutaneous administration, the carrier optionally comprises apenetration enhancing agent and/or a suitable wetting agent, optionallycombined with suitable additives of any nature in minor proportions,which additives do not cause a significant deleterious effect to theskin. Said additives may facilitate the administration to the skinand/or may be helpful for preparing the desired compositions. Thesecompositions may be administered in various ways, e.g., as a transdermalpatch, as a spot-on, as an ointment. It is especially advantageous toformulate the aforementioned pharmaceutical compositions in dosage unitform for ease of administration and uniformity of dosage. Dosage unitform as used in the specification and claims herein refers to physicallydiscrete units suitable as unitary dosages, each unit containing apredetermined quantity of active ingredient calculated to produce thedesired therapeutic effect in association with the requiredpharmaceutical carrier. Examples of such dosage unit forms are tablets(including scored or coated tablets), capsules, pills, powder packets,wafers, injectable solutions or suspensions, teaspoonfuls,tablespoonfuls and the like, and segregated multiples thereof.

It is especially advantageous to formulate the aforementionedpharmaceutical compositions in dosage unit form for ease ofadministration and uniformity of dosage. Dosage unit form as used in thespecification and claims herein refers to physically discrete unitssuitable as unitary dosages, each unit containing a predeterminedquantity of active ingredient, calculated to produce the desiredtherapeutic effect, in association with the required pharmaceuticalcarrier. Examples of such dosage unit forms are tablets (includingscored or coated tablets), capsules, pills, powder packets, wafers,injectable solutions or suspensions, teaspoonfuls, tablespoonfuls andthe like, and segregated multiples thereof.

The compound of the invention is administered in an amount sufficient toinhibit cell cycle specific kinases.

In particular, the compound of the invention is administered in anamount sufficient to inhibit CDK4, AURORA A and/or AURORA B or in anamount sufficient to modulate the interaction between CDK4, AURORA Aand/or AURORA B and other genes and/or gene products involved in thecell cycle.

Even more in particular, the compound of the invention is administeredin an amount sufficient to inhibit a proliferative disorder and/or adifferentiative disorder.

With the term “other genes and/or gene products involved in the cellcycle” is meant for example genes and gene products involved inchromatin binding, formation of replication complexes, replicationlicensing, phosphorylation or other secondary modification activity,proteolytic degradation, microtubule binding, actin binding, septinbinding, microtubule organising centre nucleation activity and bindingto components of the cell cycle (signalling) pathway.

Those skilled in the art could easily determine the effective amountfrom the test results presented hereinafter. In general it iscontemplated that a therapeutically effective amount would be from 0.005mg/kg to 100 mg/kg body weight, and in particular from 0.005 mg/kg to 10mg/kg body weight. It may be appropriate to administer the required doseas single, two, three, four or more sub-doses at appropriate intervalsthroughout the day. Said sub-doses may be formulated as unit dosageforms, for example, containing 0.5 to 500 mg, and in particular 10 mg to500 mg of active ingredient per unit dosage form.

As another aspect of the present invention, a combination of a cellcycle specific kinase inhibitor of formula (I) with another medicinalagent is envisaged, especially for use as a medicine, more specificallyin the treatment of cancer or related diseases.

For the treatment of the above conditions, the compounds of theinvention may be advantageously employed in combination with one or moreother medicinal agents, more particularly, with other anti-canceragents. Examples of anti-cancer agents are:

-   -   platinum coordination compounds for example cisplatin,        carboplatin or oxalyplatin;    -   taxane compounds for example paclitaxel or docetaxel;    -   topoisomerase I inhibitors such as camptothecin compounds for        example irinotecan or topotecan;    -   topoisomerase II inhibitors such as anti-tumour podophyllotoxin        derivatives for example etoposide or teniposide;    -   anti-tumour vinca alkaloids for example vinblastine, vincristine        or vinorelbine;    -   anti-tumour nucleoside derivatives for example 5-fluorouracil,        gemcitabine or capecitabine;    -   alkylating agents such as nitrogen mustard or nitrosourea for        example cyclophosphamide, chlorambucil, carmustine or lomustine;    -   anti-tumour anthracycline derivatives for example daunorubicin,        doxorubicin, idarubicin or mitoxantrone;    -   HER2 antibodies for example trastuzumab;    -   estrogen receptor antagonists or selective estrogen receptor        modulators for example tamoxifen, toremifene, droloxifene,        faslodex or raloxifene;    -   aromatase inhibitors such as exemestane, anastrozole, letrazole        and vorozole;    -   differentiating agents such as retinoids, vitamin D and retinoic        acid metabolism blocking agents (RAMBA) for example accutane;    -   DNA methyl transferase inhibitors for example azacytidine;    -   kinase inhibitors for example flavoperidol, imatinib mesylate or        gefitinib;    -   farnesyltransferase inhibitors;    -   HDAC inhibitors;    -   other inhibitors of the ubiquitin-proteasome pathway for example        Velcade; or    -   Yondelis.

The term “platinum coordination compound” is used herein to denote anytumour cell growth inhibiting platinum coordination compound whichprovides platinum in the form of an ion.

The term “taxane compounds” indicates a class of compounds having thetaxane ring system and related to or derived from extracts from certainspecies of yew (Taxus) trees.

The term “topisomerase inhibitors” is used to indicate enzymes that arecapable of altering DNA topology in eukaryotic cells. They are criticalfor important cellular functions and cell proliferation. There are twoclasses of topoisomerases in eukaryotic cells, namely type I and typeII. Topoisomerase I is a monomeric enzyme of approximately 100,000molecular weight. The enzyme binds to DNA and introduces a transientsingle-strand break, unwinds the double helix (or allows it to unwind)and subsequently reseals the break before dissociating from the DNAstrand. Topisomerase II has a similar mechanism of action which involvesthe induction of DNA strand breaks or the formation of free radicals.

The term “camptothecin compounds” is used to indicate compounds that arerelated to or derived from the parent camptothecin compound which is awater-insoluble alkaloid derived from the Chinese tree Camptothecinacuminata and the Indian tree Nothapodytes foetida.

The term “podophyllotoxin compounds” is used to indicate compounds thatare related to or derived from the parent podophyllotoxin, which isextracted from the mandrake plant.

The term “anti-tumour vinca alkaloids” is used to indicate compoundsthat are related to or derived from extracts of the periwinkle plant(Vinca rosea).

The term “alkylating agents” encompass a diverse group of chemicals thathave the common feature that they have the capacity to contribute, underphysiological conditions, alkyl groups to biologically vitalmacromolecules such as DNA. With most of the more important agents suchas the nitrogen mustards and the nitrosoureas, the active alkylatingmoieties are generated in vivo after complex degradative reactions, someof which are enzymatic. The most important pharmacological actions ofthe alkylating agents are those that disturb the fundamental mechanismsconcerned with cell proliferation in particular DNA synthesis and celldivision. The capacity of alkylating agents to interfere with DNAfunction and integrity in rapidly proliferating tissues provides thebasis for their therapeutic applications and for many of their toxicproperties.

The term “anti-tumour anthracycline derivatives” comprise antibioticsobtained from the fungus Strep. peuticus var. caesius and theirderivatives, characterised by having a tetracycline ring structure withan unusual sugar, daunosamine, attached by a glycosidic linkage.

Amplification of the human epidermal growth factor receptor 2 protein(HER 2) in primary breast carcinomas has been shown to correlate with apoor clinical prognosis for certain patients. Trastuzumab is a highlypurified recombinant DNA-derived humanized monoclonal IgG1 kappaantibody that binds with high affiniity and specificity to theextracellular domain of the HER2 receptor.

Many breast cancers have estrogen receptors and growth of these tumourscan be stimulated by estrogen. The terms “estrogen receptor antagonists”and “selective estrogen receptor modulators” are used to indicatecompetitive inhibitors of estradiol binding to the estrogen receptor(ER). Selective estrogen receptor modulators, when bound to the ER,induces a change in the three-dimensional shape of the receptor,modulating its binding to the estrogen responsive element (ERE) on DNA.

In postmenopausal women, the principal source of circulating estrogen isfrom conversion of adrenal and ovarian androgens (androstenedione andtestosterone) to estrogens (estrone and estradiol) by the aromataseenzyme in peripheral tissues. Estrogen deprivation through aromataseinhibition or inactivation is an effective and selective treatment forsome postmenopausal patients with hormone-dependent breast cancer.

The term “antiestrogen agent” is used herein to include not onlyestrogen receptor antagonists and selective estrogen receptor modulatorsbut also aromatase inhibitors as discussed above.

The term “differentiating agents” encompass compounds that can, invarious ways, inhibit cell proliferation and induce differentiation.Vitamin D and retinoids are known to play a major role in regulatinggrowth and differentiation of a wide variety of normal and malignantcell types. Retinoic acid metabolism blocking agents (RAMBA's) increasethe levels of endogenous retinoic acids by inhibiting the cytochromeP450-mediated catabolism of retinoic acids.

DNA methylation changes are among the most common abnormalities in humanneoplasia. Hypermethylation within the promotors of selected genes isusually associated with inactivation of the involved genes. The term“DNA methyl transferase inhibitors” is used to indicate compounds thatact through pharmacological inhibition of DNA methyl transferase andreactivation of tumour suppressor gene expression.

The term “kinase inhibitors” comprises potent inhibitors of kinases thatare involved in cell signalling, cell cycle progression and programmedcell death (apoptosis).

The term “farnesyltransferase inhibitors” is used to indicate compoundsthat were designed to prevent farnesylation of Ras and otherintracellular proteins. They have been shown to have effect on malignantcell proliferation and survival.

The term “histone deacetylase inhibitor” or “inhibitor of histonedeacetylase” is used to identify a compound, which is capable ofinteracting with a histone deacetylase and inhibiting its activity, moreparticularly its enzymatic activity. Inhibiting histone deacetylaseenzymatic activity means reducing the ability of a histone deacetylaseto remove an acetyl group from a histone.

The term “other inhibitors of the ubiquitin-proteasome pathway” is usedto identify compounds that inhibit the targeted destruction of cellularproteins in the proteasome, including cell cycle regulatory proteins.

In view of their useful pharmacological properties, the components ofthe combinations according to the invention, i.e. the other medicinalagent and a cell cycle specific kinase inhibitor of formula (I) may beformulated into various pharmaceutical forms for administrationpurposes. The components may be formulated separately in individualpharmaceutical compositions or in a unitary pharmaceutical compositioncontaining both components.

The present invention therefore also relates to a pharmaceuticalcomposition comprising the other medicinal agent and a cell cyclespecific kinase inhibitor of formula (I) together with one or morepharmaceutical carriers.

The present invention further relates to the use of a combinationaccording to the invention in the manufacture of a pharmaceuticalcomposition for inhibiting a proliferative disorder and/or adifferentiative disorder.

The present invention further relates to a product containing as firstactive ingredient a cell cycle specific kinase inhibitor of formula (I)and as second active ingredient another medicinal agent, as a combinedpreparation for simultaneous, separate or sequential use in thetreatment of patients suffering from a proliferative disorder and/or adifferentiative disorder.

The other medicinal agent and a cell cycle specific kinase inhibitor offormula (I) may be administered simultaneously (e.g. in separate orunitary compositions) or sequentially in either order. In the lattercase, the two compounds will be administered within a period and in anamount and manner that is sufficient to ensure that an advantageous orsynergistic effect is achieved. It will be appreciated that thepreferred method and order of administration and the respective dosageamounts and regimes for each component of the combination will depend onthe particular other medicinal agent and the cell cycle specific kinaseinhibitor of formula (I) being administered, their route ofadministration, the particular proliferative disorder and/ordifferentiative disorder being treated and the particular host beingtreated. The optimum method and order of administration and the dosageamounts and regime can be readily determined by those skilled in the artusing conventional methods and in view of the information set outherein.

The platinum coordination compound is advantageously administered in adosage of 1 to 500 mg per square meter (mg/m²) of body surface area, forexample 50 to 400 mg/m², particularly for cisplatin in a dosage of about75 mg/m² and for carboplatin in about 300 mg/m² per course of treatment.

The taxane compound is advantageously administered in a dosage of 50 to400 mg per square meter (mg/m²) of body surface area, for example 75 to250 mg/m², particularly for paclitaxel in a dosage of about 175 to 250mg/m² and for docetaxel in about 75 to 150 mg/m² per course oftreatment.

The camptothecin compound is advantageously administered in a dosage of0. 1 to 400 mg per square meter (mg/m²) of body surface area, forexample 1 to 300 mg/m², particularly for irinotecan in a dosage of about100 to 350 mg/m² and for topotecan in about 1 to 2 mg/m² per course oftreatment.

The anti-tumour podophyllotoxin derivative is advantageouslyadministered in a dosage of 30 to 300 mg per square meter (mg/m²) ofbody surface area, for example 50 to 250 mg/m², particularly foretoposide in a dosage of about 35 to 100 mg/m² and for teniposide inabout 50 to 250 mg/m² per course of treatment.

The anti-tumour vinca alkaloid is advantageously administered in adosage of 2 to 30 mg per square meter (mg/m²) of body surface area,particularly for vinblastine in a dosage of about 3 to 12 mg/m², forvincristine in a dosage of about 1 to 2 mg/m², and for vinorelbine indosage of about 10 to 30 mg/m² per course of treatment.

The anti-tumour nucleoside derivative is advantageously administered ina dosage of 200 to 2500 mg per square meter (mg/m²) of body surfacearea, for example 700 to 1500 mg/m², particularly for 5-FU in a dosageof 200 to 500 mg/m², for gemcitabine in a dosage of about 800 to 1200mg/m² and for capecitabine in about 1000 to 2500 mg/m² per course oftreatment.

The alkylating agents such as nitrogen mustard or nitrosourea isadvantageously administered in a dosage of 100 to 500 mg per squaremeter (mg/m²) of body surface area, for example 120 to 200 mg/m²,particularly for cyclophosphamide in a dosage of about 100 to 500 mg/m²,for chlorambucil in a dosage of about 0.1 to 0.2 mg/kg, for carmustinein a dosage of about 150 to 200 mg/m², and for lomustine in a dosage ofabout 100 to 150 mg/m² per course of treatment.

The anti-tumour anthracycline derivative is advantageously administeredin a dosage of 10 to 75 mg per square meter (mg/m²) of body surfacearea, for example 15 to 60 mg/m², particularly for doxorubicin in adosage of about 40 to 75 mg/m², for daunorubicin in a dosage of about 25to 45mg/m², and for idarubicin in a dosage of about 10 to 15 mg/m² percourse of treatment.

Trastuzumab is advantageously administered in a dosage of 1 to 5 mg persquare meter (mg/m²) of body surface area, particularly 2 to 4 mg/m² percourse of treatment.

The antiestrogen agent is advantageously administered in a dosage ofabout 1 to 100 mg daily depending on the particular agent and thecondition being treated. Tamoxifen is advantageously administered orallyin a dosage of 5 to 50 mg, preferably 10 to 20 mg twice a day,continuing the therapy for sufficient time to achieve and maintain atherapeutic effect. Toremifene is advantageously administered orally ina dosage of about 60 mg once a day, continuing the therapy forsufficient time to achieve and maintain a therapeutic effect.Anastrozole is advantageously administered orally in a dosage of about 1mg once a day. Droloxifene is advantageously administered orally in adosage of about 20-100 mg once a day. Raloxifene is advantageouslyadministered orally in a dosage of about 60 mg once a day. Exemestane isadvantageously administered orally in a dosage of about 25 mg once aday.

These dosages may be administered for example once, twice or more percourse of treatment, which may be repeated for example every 7, 14, 21or 28 days.

The compounds of formula (I), the acid addition salts and stereoisomericforms thereof can have valuable diagnostic properties in that they canbe used for detecting or identifying an a cell cycle specific kinase ina biological sample comprising detecting or measuring the formation of acomplex between a labelled compound and/or CDK4, AURORA A and/or AURORAB and/or other molecules, peptides, proteins, enzymes or receptors.

The detecting or identifying methods can use compounds that are labelledwith labelling agents such as radioisotopes, enzymes, fluorescentsubstances, luminous substances, etc. Examples of the radioisotopesinclude ¹²⁵I, ¹³¹I, ³H and ¹⁴C. Enzymes are usually made detectable byconjugation of an appropriate substrate which, in turn catalyses adetectable reaction. Examples thereof include, for example,beta-galactosidase, beta-glucosidase, alkaline phosphatase, peroxidaseand malate dehydrogenase, preferably horseradish peroxidase. Theluminous substances include, for example, luminol, luminol derivatives,luciferin, aequorin and luciferase. Biological samples can be defined asbody tissue or body fluids. Examples of body fluids are cerebrospinalfluid, blood, plasma, serum, urine, sputum, saliva and the like.

Experimental Part

Hereinafter, the term ‘THF’ means tetrahydrofuran, ‘EtOH’ means ethanol,‘DMSO’ means dimethylsulfoxide, ‘DCM’ means dichloromethane, ‘DIPE’means diisopropyl ether, ‘EtOAc’ means ethyl acetate, ‘Et₂O’ meansdiethyl ether, ‘HBTU’ means1-[bis(dimethylamino)methylene]-1H-benzotriazolium,,hexafluorophosphate(1-), 3-oxide, ‘HOBt’ means1-hydroxy-1H-benzotriazole, ‘LiAlH₄’ means lithiumaluminiumhydride,‘MeOH’ means methanol, ‘mcPBA’ means 3-chlorobenzenecarboperoxoic acid.‘TEA’ means triethylamine and ‘TFA’ means trifluoroacetic acid.

A. Preparation of the Intermediate Compounds EXAMPLE A1

a) Preparation of Intermediate 1

TEA (0.159 mol) was added at room temperature to a mixture of4-chloro-2-(methylthio)-5-pyrimidinecarboxylic acid ethyl ester (0.0534mol) and 2-cyclohexylhydrazinecarboxylic acid 1,1-dimethylethyl ester(0.1068 mol) in THF (125 ml). The mixture was stirred at roomtemperature overnight. The solvent was evaporated. The residue was takenup in DCM. The organic layer was washed with saturated NaHCO₃, dried(MgSO₄), filtered and the solvent was evaporated, yielding 37.2 g(>100%) of intermediate 1.b) Preparation of Intermediate-2

A solution of intermediate 1 (0.0633 mol) in THF (200 ml) was addeddropwise at room temperature to a suspension of LiAlH₄ (0.1014 mol) inTHF (200 ml). The mixture was stirred at room temperature for 3 hours.EtOAc was added dropwise. Then water (6 ml) was added. The mixture wasfiltered over celite. Celite was washed with EtOAc. The filtrate wasevaporated. The residue (23 g) was purified by column chromatographyover silica gel (35-70 μm) (eluent: DCM/MeOH/NH₄OH 96/4/0.5). Thedesired fractions were collected and the solvent was evaporated,yielding 2.2 g (9%) of intermediate 2.c) Preparation of Intermediate 3

Manganese dioxide (2.7 g) was added at room temperature to a mixture ofintermediate 2 (0.0057 mol) in DCM (100 ml). The mixture was stirred atroom temperature for 24 hours. MnO₂ (1 g) was added. The mixture wasstirred for 24 hours, then filtered over celite. The filtrate wasevaporated, yielding: 2.2 g (>100%) of intermediate 3.d) Preparation of Intermediate 4

HCl/2-propanol (2.2 ml) was added at room temperature to a mixture ofintermediate 3 (0.006 mol) in EtOH (20 ml). The mixture was stirred atroom temperature for 5 hours. The solvent was evaporated. The residuewas taken up into ice water. The aqueous layer was basified with K₂CO₃.The mixture was extracted with DCM. The organic layer was separated,dried (MgSO₄), filtered and the solvent was evaporated, yielding 1.3 g(88%) of intermediate 4.e) Preparation of Intermediated 5

mcPBA 70% (0.0104 mol) was added at room temperature to a mixture ofintermediate 4 (0.0052 mol) in DCM (30 ml). The mixture was stirred atroom temperature overnight. K₂CO₃ 10% was added. The mixture wasextracted with DCM. The organic layer was separated, dried (MgSO₄),filtered and the solvent was evaporated, yielding 1.35 g (90%) ofintermediate 5.

EXAMPLE A2

a) Preparation of Intermediate 6

In a 100-ml two-neck reaction flask,4-[[[(1,1-dimethylethyl)dimethylsilyl]oxy]methyl]-benzenamine (0.01662mol) was dissolved in THF dry (25 ml). The solution was heated to 60° C.on an oil-bath. A solution of dicarbonic acid, bis(1,1-dimethylethyl)ester (0.02078 mol) in THF dry (15 ml) was added dropwise and thereaction mixture was stirred for ±4 hours. The solvent was evaporated.The residue (6.378 g) was purified by column chromatography over silicagel (eluent: EtOAc/hexane 5/95), then repurified by columnchromatography over silica gel (eluent: DCM/hexane 1/1, then 6/4, then7/3, ending with 100% DCM). The product fractions were collected and thesolvent was evaporated, yielding 3.625 g (65%) of intermediate 6.b) Preparation of Intermediate 7

Intermediate 6 (0.00892 mol) was kept for 15 min in vacuo, then Argonwas let in. Reaction was done under Argon. Intermediate 6 was dissolvedin THF dry (15 ml) and the solution was cooled to 0° C. (15 min).Ethylmagnesium chloride 2.8M/THF (0.00446 mol) was added dropwise andthe mixture was stirred for 2 hours at 0° C. to give solution I.Starting material intermediate 5 was kept for 15 min in vacuo, thenArgon was let in. Intermediate 5 was dissolved in THF dry (15 ml) andthis solution was added dropwise to the solution I. The reaction mixturewas stirred until reaction was complete (ca. 40 minutes). The mixturewas extracted with Et₂O/saturated aqueous NaHCO₃ solution/saturatedaqueous NaCl solution. The separated organic layer was dried (Na₂SO₄),filtered and the solvent evaporated. The residue (3.350 g) was purifiedby column chromatography over silica gel (eluent: EtOAc/hexane 3/7). Theproduct fractions were collected and the solvent was evaporated,yielding 0.921 g (96%) of intermediate 7.c) Preparation of Intermediate 8

Intermediate 7 (0.000726 mol) was dissolved in THF dry (3 ml). Asolution of tetrabutylammonium fluoride trihydrate (0.000871 mol) in THFdry (2 ml) was added dropwise. The reaction mixture was stirred at roomtemperature for one hour. This mixture was extracted with Et₂O/water(2×), then with an aqueous NaCl solution. The extract was filtered andthe solvent was evaporated. The residue (0.400 g) was purified by columnchromatography over silica gel (eluent: DCM/EtOAc/toluene 10/10/0.5).Two product fraction groups were collected. The purest fractions werecombined and the solvent was evaporated, yielding 0.225 g (73%) ofintermediate 8.d) Preparation of Intermediate 9

Manganese dioxide (0.011075 mol) was stirred in DCM (6 ml). Theresulting suspension was stirred for 15 min at 0° C. A solution ofintermediate 8 (0.000443 mol) in DCM (6 ml) was added dropwise and thereaction mixture was stirred for 2.5 hours at 0° C. The mixture wasfiltered over Na₂SO₄ and the filter cake was rinsed with DCM, then withEtOAc. The filtrate was evaporated, yielding 0.172 g of intermediate 9(light foam, used in next reaction step, without further purification).e) Preparation of Intermediate 10

A solution of intermediate 9 (100 mg, 0.237 mmol) in dry THF (2 ml) wastreated with 4-(2-aminoethyl)morpholine (62 ml, 0.475 mmol) and aceticacid (27 ml) and stirred at room temperature for 2 hours. After theaddition of NaBH(OAc)₃ (151 mg, 0.712 mmol) stirring was continued for27 hours. Normal workup (EtOAc, saturated aqueous NaHCO₃ solution,saturated aqueous NaCl solution, Na₂SO₄) and flash chromatography(DCM/MeOH 100:0 to 80:20), yielding 109 mg (85.7%) of intermediate 10.

EXAMPLE A3

a) Preparation of Intermediate 11

A solution of cyclohexanamine (0.062 mol) in N,N-dimethylacetamide (20ml) was added dropwise to a cooled (−10° C.) solution of2,4-dichloro-5-nitropyrimidine (0.062 mol) and DIPE (8.1 g) inN,N-dimethylacetamide (80 ml), then the reaction mixture was allowed toreach room temperature overnight, yielding intermediate 11 (containsregio-isomer; used as such in the next reaction steps).b) Preparation of Intermediate 12

A mixture of intermediate 11 (0.031 mol),4-(4-methyl-1-piperazinyl)-benzenamine, hydrochloride (0.031 mol) andDIPE (10 g) was heated at 60° C. for 3 hours, then the reaction mixturewas cooled and added dropwise to water (200 ml). The resulting solidswere filtered off and dried in a vacuo at 60° C., yielding 9.6 g ofintermediate 12.c) Preparation of Intermediate 13

A mixture of intermediate 12 (0.023 mol) and TEA (10 ml) in THF (250 ml)was hydrogenated at 50° C. with Pd/C 10% (2 g) as a catalyst in thepresence of thiophene solution (1 ml). After uptake of H₂ (3 equiv.),the catalyst was filtered off and the filtrate was evaporated. Theresidue was dissolved in DCM, washed with water and dried, yielding 6.7g (76.5%) of intermediate 13.

EXAMPLE A4

a) Preparation of Intermediate 14

HCl/2-propanol (18 ml) was added to a mixture of intermediate 1 (0.0292mol) in EtOH (120 ml). The mixture was stirred and refluxed for 3 hoursand 30 minutes, then stirred at room temperature overnight. The solventwas evaporated. The residue was taken up in DIPE. The precipitate wasfiltered off and dried, yielding 6.5 g of intermediate 14, isolated as ahydrochloric acid salt. This fraction was used directly in the nextreaction step.b) Preparation of Intermediate 15

mcPBA 70% (0.0432 mol) was added portionwise at room temperature to amixture of intermediate 14 (0.0216 mol) in DCM (80 ml). The mixture wasstirred at room temperature overnight. The mixture was purified bycolumn chromatography over silica gel (70-200 μm) (eluent: DCMNMeOH95/5). The pure fractions were collected and the solvent was evaporated,yielding 2.2 g (36%) of intermediate 15.

EXAMPLE A5

a) Preparation of Intermediate 16

Reaction in a sealed tube. A suspension of intermediate 5 (0.00178 mol),4-amino-benzoic acid, 1,1-dimethylethyl ester (0.00267 mol) and cesiumcarbonate (0.00267 mol) in DMSO, abs. (5 ml) was stirred for 2 hours at100° C. This mixture was extracted with EtOAc (3×), water. The aqueousphase was treated with 1 N HCl until pH=5. The acidic layer wasre-extracted with EtOAc. The extract was washed with a saturated aqueousNaCl solution, then dried (Na₂SO₄), filtered and the solvent wasevaporated. The residue was purified by column chromatography oversilica gel (eluent: DCM/hexane 9/1, over 100% DCM, to hexane/EtOAc 1/1).The product fractions were collected and the solvent was evaporated,yielding 0.445 g (63.52%) of intermediate 16.b) Preparation of Intermediate 17

Intermediate 16 (0.001105 mol) was dissolved in DCM (10 ml). TFA(I0 ml)was added dropwise via an addition funnel. The reaction mixture wasstirred for one hour at room temperature. The solvent was evaporated.Toluene was added and azeotroped on the rotary evaporator (2×), yielding0.564 g of intermediate 17.

B. Preparation of the Final Compounds EXAMPLE B1

Preparation of Compounds 1

A mixture of intermediate 5 (0.0026 mol) and4-(4-methyl-1-piperazinyl)-benzenamine (0.0107 mol) was stirred at 140°C. for 1 hour, then brought to room temperature. DCM was added. Theresidue (3.3g) was purified by column chromatography over silica gel(15-35 μm) (eluent: DCM/MeOH/NH₄OH 94/6/0.1). The pure fractions werecollected and the solvent was evaporated. The residue (0.72 g) was takenup in Et₂O. The precipitate was filtered off and dried, yielding 0.51 g(49%) of compound 1, melting point: 180° C.

EXAMPLE B2

Preparation of Compound 2

Reaction under Ar flow. Dry DMSO (0.25 ml) was added to intermediate 5(0.000214 mol) and 4-[(4-methyl-1-piperazinyl)methyl]-benzenamine(0.000321 mol), under Argon. The reaction mixture was stirred for ±20hours at 100° C. (oil-bath). This mixture was extracted withCHCl₃/2×NaHCO₃/2× water. The extract was filtered over silica gel(MgSO₄) and the solvent was evaporated. The residue (0.097 g) waspurified by preperative HPLC (eluent: DCM/MeOH). The product fractionswere collected and the solvent was evaporated, yielding 0.030 g (oil) ofcompound 2.

EXAMPLE B3

Preparation of Compound-3

Reaction under Ar flow. Dry DMSO (1.5 ml) was added to a mixture ofintermediate 5 (0.000428 mol), 1-(4-aminobenzoyl)-4-methyl-piperazine(0.000642 mol) and cesium carbonate (0.000642 mol). The reaction mixturewas stirred for 3 hours at 100° C. This mixture was extracted with amixture of EtOAc/NaHCO₃/H₂O/NaCl. The extract's solvent was evaporated.The residue (0.145 g) was purified by column chromatography over silicagel (eluent: DCM/MeOH gradient). The product fractions were collectedand the solvent was evaporated, yielding 0.094 g of compound 3.

EXAMPLE B4

Preparation of Compound 4

TFA (1.5 ml) was added to a solution of intermediate 10 (103 mg, 0.192mmol) in dry DCM (1.5 ml). The solution was stirred at room temperaturefor 60 minutes followed by a normal workup (EtOAc, saturated aqueousNaHCO₃ solution, saturated aqueous NaCl solution, Na₂SO₄) and flashchromatography (DCM/MeOH 100:0 to 70:30), yielding 73 mg (87%) ofcompound 4 as colorless foam.

EXAMPLE B5

Preparation of Compound 5

A mixture of intermediate 13 (0.00102 mol) and Gold's reagent (0.00144mol) in dioxane (10 ml) was stirred and refluxed overnight, then thesolvent was evaporated and the residue was partitioned betweenDCM/water. The layers were separated and the aqueous layer was extractedwith DCM. The organic layers were combined, dried (MgSO₄), filtered offand the solvent was evaporated. The residue (0.43 g) was purified byreversed phase high-performance liquid chromatography. The purefractions were collected and the solvent was evaporated. The residue wasextracted with DCM (3×30 ml). The organic layer was dried (MgSO₄),filtered off and the solvent was evaporated, yielding 0.200 g ofcompound 5, melting point: 203.5-203.6° C.

EXAMPLE B6

Preparation of Compound 6

A mixture of intermediate 13 (0.00144 mol) and urea (0.002 mol) inxylene (50 ml) was stirred and refluxed for 16 hours. The solvent wasevaporated and the residue was purified by reversed phasehigh-performance liquid chromatography. The product fractions werecollected and the solvent was evaporated, yielding 0.0767 g of compound6, melting point: 294° C.

EXAMPLE B7

Preparation of Compound 7

A mixture of intermediate 15 (0.0035 mol) and4-(4-methyl-1-piperazinyl)-benzenamine (0.0142 mol) was stirred at 140°C. for 2 hours, then brought to room temperature. DCM was added. Theprecipitate was filtered, washed several times with EtOH and dried withEt₂O, yielding 0.244 g (17%) of compound 7, melting point: >260° C.

EXAMPLE B8

Preparation of Compound 8

A mixture of intermediate 5 (80 mg, 0.285 mmol),4-(morpholinosulfonyl)aniline (76 mg, 0.314 mmol) and cesium carbonate(102 mg, 0.314 mmol) was heated in dry DMSO (1 ml) to 100° C. for 12hours. Treatment of the mixture with EtOAc and saturated aqueous NaHCO₃solution resulted in formation of a precipitate in the organic layer.The organic layer was separated, washed (water, saturated aqueous NaClsolution) and centrifuged. The pellet was washed (Et₂O/MeOH, Et₂O) anddried in vacuo, yielding 51 mg (40.5%) of compound 8, melting point 240°C.

EXAMPLE B9

Preparation of Compound 9

A suspension of intermediate 17 (0.000207 mol),4-amino-1-piperidineethanol, dihydrochloride (0.000311 mol), HOBt(0.0003111 mol) and HBTU (0.0003111 mol) in DMF dry (1.5 ml) was stirredat room temperature. DIPE (0.001035 mol) was added and the reactionmixture was stirred overnight at room temperature. This mixture wasextracted with Et₂O/aqueous NaHCO₃ solution, followed by water (2×). Theseparated organic layer was dried (Na₂SO₄), filtered and the solvent wasevaporated. The residue (0.090 g) was purified by column chromatographyover silica gel (eluent: DCM/MeOH 93/7, then 9/1; then eluent:DCM/(MeOH/NH₃ 1%) 9/1; then: DCM/(MeOH/NH₃ 2%) 9/1). The productfractions were collected and the solvent was evaporated, yielding 0.052g of compound 9, melting point 177-181° C.

Table F-1 lists the compounds that were prepared according to the abovedescribed synthesis schemes. TABLE F-1

Co. No. 1; Ex. [B1]; mp. 180° C.

Co. No. 2; Ex. [B2]

Co. No. 3; Ex. [B3]

Co. No. 4; Ex. [B4]

Co. No. 5; Ex. [B5]; mp. 203.5-203.6° C.

Co. No. 6; Ex. [B6]; mp. 294° C.

Co. No. 7; Ex. [B7]; mp. >260° C.

Co. No. 8; Ex. [B8]; mp. 240° C.

Co. No. 9; Ex. [B9]; mp. 177-181° C.

Co. No. 10; Ex. [B1]

Co. No. 11; Ex. [B1]

Co. No. 12; Ex. [B1]

Co. No. 13; Ex. [B1]

Co. No. 14; Ex. [B1]

Co. No. 15; Ex. [B1]

(ENDO); Co. No. 16; Ex. [B1]

Co. No. 17; Ex. [B1]; mp. 183.5-185° C.

Co. No. 18; Ex. [B1]; mp 170-172° C.

Co. No. 19; Ex. [B1]; mp. 194-196.5° C.

Co. No. 20; Ex. [B1]

Co. No. 21; Ex. [B1]

Co. No. 22; Ex. [B1]; mp. 157-163° C.

(EXO); Co. No. 23; Ex. [B1]; mp. 192.5-193.5° C.

Co. No. 24; Ex. [B1]; mp. 150.5-152° C.

Co. No. 25; Ex. [B1]; mp. 159-161° C.

Co. No. 26; Ex. [B1]

Co. No. 27; Ex. [B1]; mp. 196-198° C.

Co. No. 28; Ex. [B1]; mp. 228-230° C.

Co. No. 29; Ex. [B1]; mp. 194-196.5° C.

Co. No. 30; Ex. [B1]; mp. 204-206° C.

Co. No. 31; Ex. [B1]

Co. No. 32; Ex. [B1]

Co. No. 33; Ex. [B2]; mp. 135-137° C.

.C₂HF₃O₂; Co. No. 34; Ex. [B2]

Co. No. 35; Ex. [B2]

(ENDO); Co. No. 36; Ex. [B2]

Co. No. 37; Ex. [B2]

Co. No. 38; Ex. [B2]

(EXO); Co. No. 39; Ex. [B2]

(EXO); Co. No. 40; Ex. [B2]

(EXO); Co. No. 41; Ex. [B2]

Co. No. 42; Ex. [B2]

Co. No. 43; Ex. [B2]

Co. No. 44; Ex. [B2]

Co. No. 45; Ex. [B2]; mp. 134.5-137° C.

Co. No. 46; Ex. [B2]

Co. No. 47; Ex. [B2]

Co. No. 48; Ex. [B2]

Co. No. 49; Ex. [B2]

Co. No. 50; Ex. [B2]

Co. No. 51; Ex. [B2]; mp. 161.5-165° C.

(EXO); Co. No. 52; Ex. [B2]; mp. 161.5-164° C.

Co. No. 53; Ex. [B2]

Co. No. 54; Ex. [B2]

Co. No. 55; Ex. [B2]

Co. No. 56; Ex. [B3]; mp. 207-208.5° C.

.C2HF3O2; (ENDO); Co. No. 57; Ex. [B3]

Co. No. 58; Ex. [B3]

.2HCl; Co. No. 59; Ex. [B4]; mp. 249-250° C.

.2HCl; Co. No. 60; Ex. [B4]; mp. 154-156° C.

Co. No. 61; Ex. [B4]

Co. No. 62; Ex. [B4]

Co. No. 63; Ex. [B4]

(EXO); Co. No. 64; Ex. [B4]

(EXO); Co. No. 65; Ex. [B4]

Co. No. 66; Ex. [B4]

Co. No. 67; Ex. [B4]

(EXO); Co. No. 68; Ex. [B4]

(EXO); Co. No. 69; Ex. [B4]

Co. No. 70; Ex. [B4]

Co. No. 71; Ex. [B4]

Co. No. 72; Ex. [B4]

Co. No. 73; Ex. [B4]

Co. No. 74; Ex. [B4]

Co. No. 75; Ex. [B4]

Co. No. 76; Ex. [B4]

Co. No. 77; Ex. [B4]

Co. No. 78; Ex. [B6]

Co. No. 79; Ex. [B6]; mp. 230-232° C.

Co. No. 80; Ex. [B6]; mp. 248-252° C.

Co. No. 81; Ex. [B6]; mp. >250° C.

Co. No. 82; Ex. [B6]

Co. No. 83; Ex. [B6]; mp. 240-243° C.

Co. No. 84; Ex. [B6]; mp. 248-251° C.

Co. No. 85; Ex. [B6]; mp. >250° C.

.2 HCl; Co. No. 86; Ex. [B6]; mp. >250° C.

.2 HCl; Co. No. 87; Ex. [B6]; mp. >250° C.

(EXO); Co. No. 88; Ex. [B6]; mp. >250° C.

Co. No. 89; Ex. [B6]

Co. No. 90; Ex. [B6]; mp. 173-174° C.

Co. No. 91; Ex. [B7]

Co. No. 92; Ex. [B7]

Co. No. 93, Ex. [B7]

Co. No. 94; Ex. [B7]

Co. No. 95; Ex. [B9]; mp. 232-233° C.

Co. No. 96; Ex. [B9]; mp. 183-185° C.

Co. No. 97; Ex. [B9]; mp. >180° C. (dec.)

Co. No. 98; Ex. [B9]; mp. 233-236° C.

C. Pharmacological Example

The pharmacological activity of the present compounds was examined usingthe following test.

C.1. In Vitro Filtration Assay for CDK4 Inhibitory Activity

Compounds of the present invention were tested in an in vitro filtrationassay assessing CDK4 activity by means of its pRb-phosphorylationactivity using [³³P]-ATP as phosphor donor. The radioactivephosphorylated pRb is then captured on filtermats and the incorporated[³³P] quantitated using a phosphorage storage screen.

The CDK4 kinase reaction is performed at 25° C. for 45 minutes in a96-well microtiterplate. The 25 μl reaction volume contains 50 mM HepespH 7.5, 10 mM NaF, 10 mM MgCl₂, 1 mM Na₃VO₄, 1 μM unlabeled ATP, 1 mMDTT, 0.5 μCi AT³³P, 0.76 μg/well GST-pRb, 50 ng CDK4/cyclinD1/well and0.2% compound in 100% DMSO.

The reaction is stopped by adding 5 μl of a 3% phosphoric acid solution.10 μl of the reaction mixture is then spotted onto a Filtermat P30filter (Wallac) and washed 3 times for 5 min. in 75 mM phosphoric acidand 1 time for 5 min. in methanol prior to drying and quantification onthe Typhoon (Amersham) using a phosphorage storage screen.

C.2. In Vitro Filtration Assay for AURORA A Inhibitory Activity

Compounds of the present invention were tested in an in vitro filtrationassay assessing AURORA A activity by means of itssubstrate-phosphorylation activity using [³³P]-ATP as phosphor donor.The radioactive phosphorylated substrate is then captured on filtermatsand the incorporated [³³P] quantitated using a phosphorage storagescreen.

The Aurora-A kinase reaction is performed at 25° C. for 40 minutes in a96-well microtiterplate. The 25 μl reaction volume contains 12 mM MOPSpH 7, 0.4 mM EDTA, 0.002% Brij35, 1% glycerol, 0.02%beta-mercapto-ethanol, 0.2 mg/ml BSA, 1 μM unlabeled ATP, 0.2 pCi[³³P]-ATP, 200 μM Kemptide, 3 ng Aurora A/well and 0.2% compound in 100%DMSO.

The reaction is stopped by adding 5 μl of a 3% phosphoric acid solution.10 μl of the reaction mixture is then spotted onto a Filtermat P30filter (Wallac) and washed 3 times for 5 min. in 75 mM phosphoric acidand 1 time for 5 min. in methanol prior to drying and quantification onthe Typhoon (Amersham) using a phosphorage storage screen.

C.3 In Vitro Filtration Assay for AURORA B Inhibitory Activity

Compounds of the present invention were tested in an in vitro filtrationassay assessing AURORA B activity by means of itssubstrate-phosphorylation activity using [³³P]-ATP as phosphor donor.The radioactive phosphorylated substrate is then captured on filtermatsand the incorporated [³³P] quantitated using a phosphorage storagescreen.

The Aurora-B kinase reaction is performed at 25° C. for 40 minutes in a96-well microtiterplate. The 25 μl reaction volume contains 60 mM HepespH 7.5, 3 mM MgCl₂, 3 mM MnCl₂, 3 μM Na₃VO₄, 50 μg/ml PEG20000, 1 μMunlabeled ATP, 1 mM DTT, 0.2 μCi AT³³P, 0.25 μg/well peptide(C(LRRWSLG)×4),100 ng Aurora-B/well and 0.2% compound in 100% DMSO. Thereaction is stopped by adding 5 μl of a 3% phosphoric acid solution. 10μl of the reaction mixture is then spotted onto a Filtermat P30 filter(Wallac) and washed 3 times for 5 min. in 75 mM phosphoric acid and 1time for 5 min. in methanol prior to drying and quantification on theTyphoon (Amersham) using a phosphorage storage screen.

C.4. In Vitro Scintillation Proximity Assay (SPA) for CDK4 InhibitoryActivity

Compounds of the present invention were tested in an in vitro assaybased on SPA technology.

In principle, the assay relies upon the well established SPA technologyfor the detection of CDK4 phosphorylated proteins, i.e pRb. Thisphosphorylation is performed using I0 CDK4/cyclinD1 enzyme complex and³³P-ATP as phosphor donor.

The CDK4 SPA kinase reaction is performed at 25° C. for 30 minutes in a96-well microtiter plate. The 100 μl reaction volume contains 40 mMHepes, 6 mM NaF, 6 mM MgCl₂, 0.6 mM Na₃VO₄, 2 μg/ml Pepstatin, 2 μg/mlLeupeptin, 2 μg/ml Aprotinin, 50 μg/ml TLCK, 150 μg/ml DTT, 0.3 μg/mlbenzamide, 0.1 μCi ³³P-ATP, 1.7 μg GST-pRb, 50 ng CDK4/cyclinD1/well and0.2% compound in 100% DMSO. The reaction is stopped by adding to eachwell 100 μl of the glutathione coated SPA beads (10 mg/ml in PBS+10 mMEDTA+100 μM ATP+0.05% Triton X 100). The plates are then shaked at 300rpm for 30 min to allow binding of the GST-tagged substrate to theglutathione coated beads. The beads are allowed to settle at the bottomof the plate for 30 minutes. The microtiterplates are centrifuges at 800rpm for 10 minutes and the amount of phosphorylated (³³P) substrate isdetermined by counting (30 sec/well) in a microtiterplate scintillationcounter.

C.5. Calculation of pIC₅₀ Values

For each experiment, controls (containing enzyme (complex) and DMSOwithout compound), a blank incubation (containing DMSO but no enzyme(complex) or compound) and samples (containing enzyme (complex) andcompound dissolved in DMSO) were run in parallel. All compounds testedwere dissolved and eventually further diluted in DMSO. In firstinstance, compounds were tested at a concentration of 10⁻⁵ M. When thecompounds showed activity at 10⁻⁵ M, a dose-response curve was madewherein the compounds were tested at concentrations between 10⁻⁵M and10⁻⁸M. In each test, the blank value was subtracted from both thecontrol and the sample values. The control sample represented maximalenzyme activity. For each sample, the amount of cpm was expressed as apercentage of the mean cpm value of the controls. When appropriate,IC₅₀-values (concentration of the drug, needed to reduce the enzymeactivity to 50% of the control) were computed using linear interpolationbetween the experimental points just above and below the 50% level.Herein the effects of test compounds are expressed as pIC₅₀ (thenegative log value of the IC₅₀-value). The inhibitory activity of thetested compounds of the invention is shown in Table-2 TABLE F-2 TableF-2 lists the results of the compounds that were tested according toexample C.1, C.2, C.3 and C.4 cdk4 SPA cdk4 filter Aurora A Aurora B Co.No. see C.4 see C.1 see C.2 see C.3 1 8.3 8.5 6.9 6.6 2 8.5 6.3 <5.0 38.2 6.3 <5.0 4 8.6 6.0 <5.8 5 5.5 6.2 <5.0 <5.0 6 7.0 6.6 <5.0 <5.0 76.8 7.1 6.4 8 7.0 6.3 <5.0 9 6.9 6.6 6.1 10 7.9 7.1 7.3 <5.0 11 8.2 7.56.8 6.4 12 7.7 7.4 7.2 6.6 13 7.3 7.2 7.3 <5.0 14 8.4 7.7 7.1 6.1 15 8.06.2 <5.0 16 7.5 6.4 <5.0 17 7.7 6.1 <5.0 18 6.6 6.8 <5.0 19 5.3 <5.0<5.0 20 7.9 6.3 <5.0 21 7.4 6.7 5.5 22 6.3 7.4 <5.0 23 8.3 7.0 6.8 246.9 6.6 5.5 25 6.0 7.0 5.5 26 7.8 6.6 5.9 27 6.7 6.3 <5.0 28 6.0 5.6<5.0 29 5.2 <5.0 <5.0 30 7.3 <5.0 <5.0 31 7.5 33 6.2 7.8 5.1 34 7.2 6.8<5.0 35 7.0 6.6 <5.0 36 7.0 6.6 <5.0 37 8.1 6.7 6.4 38 7.9 6.0 5.7 395.8 5.2 <5.0 40 7.2 6.5 6.4 41 5.4 5.5 5.5 42 <5.0 <5.0 <5.0 43 7.2 6.95.4 44 <5.0 <5.0 <5.0 45 6.4 6.7 5.8 46 <5.0 <5.0 <5.0 47 7.5 6.5 5.4 48<5.0 <5.0 <5.0 49 5.9 5.9 5.0 50 <5.0 <5.0 <5.0 51 5.3 6.0 <5.0 52 7.16.8 6.5 53 6.3 <5.0 5.1 54 6.2 <5.0 <5.0 55 <5.0 <5.0 <5.0 56 7.6 6.95.5 57 7.0 7.0 5.1 58 6.2 6.9 5.9 59 7.8 6.1 <5.0 60 8.1 5.8 <5.0 61<5.0 5.9 <5.0 62 7.6 6.8 5.2 63 7.6 6.0 5.6 64 5.2 <5.0 <5.0 65 6.7 6.36.3 66 5.1 5.0 <5.0 67 6.6 5.8 6.0 68 6.0 5.2 5.1 69 7.6 6.2 6.0 70 <5.0<5.0 <5.0 71 5.6 5.3 <5.0 72 <5.0 <5.0 <5.0 73 6.2 <5.0 <5.0 75 6.9 5.85.9 76 <5.0 <5.0 <5.0 77 7.1 <5.0 5.7 78 6.2 6.0 79 5.7 <5.0 <5.0 80 5.45.6 <5.0 81 5.3 <5.0 <5.0 82 5.7 <5.0 <5.0 83 5.6 <5.0 <5.0 84 6.2 5.2<5.0 85 <5.0 5.5 <5.0 86 5.6 5.3 <5.0 87 5.4 <5.0 <5.0 88 5.7 5.8 <5.089 6.5 5.8 5.5 90 6.5 5.8 5.8 91 5.9 6.2 5.1 <5.0 92 5.6 5.2 <5.0 93 5.65.3 5.8 <5.0 94 6.0 5.8 5.5 <5.0 95 6.1 6.9 6.2 96 6.2 7.0 5.9 97 6.26.4 6.3 98 6.4 6.5 5.8

The compounds were further evaluated on in vitro assays measuringinhibition of different kinase activities, on cell lines and eventuallyin in vivo tests.

C.6. Analytical Data The mass of the compounds was recorded with LCMS(liquid chromatography mass spectrometry). The analytical HPLC (column:Develosil RPAq 4.6×50 mm) was performed with different gradient eluentsystems at a flow rate of 1.5 ml/min with UV detection at 220 nm and 254nm. Different eluent systems were used which are described below. Thedata are gathered in Table F-3 below.

-   -   System A: 5% acetonitrile, 95% water (0.1% trifluoro acetic        acid) to 100% acetonitrile in 5 min    -   System B: 10% acetonitrile, 90% water (0.1% trifluoro acetic        acid) to 100% acetonitrile in 5 min    -   System C: 20% acetonitrile, 80% water (0.1% trifluoro acetic        acid) to 100% acetonitrile in 5 min    -   System D: 30% acetonitrile, 70% water (0.1% trifluoro acetic        acid) to 100% acetonitrile in 5 min    -   System E: 40% acetonitrile, 60% water (0.1% trifluoro acetic        acid) to 100% acetonitrile in 5 min    -   System F: acetonitrile, 50% water (0.1% trifluoro acetic acid)        to 100% acetonitrile in 5 min    -   System G: acetonitrile, 90% water (0.1% trifluoro acetic acid)        to 30% acetonitrile, 70% water (0.1% trifluoro acetic acid) in 5        min    -   System H: 10% acetronitrile, 90% water (0.1% trifluoro acetic        acid) to 40% acetonitrile, 60% water (0.1% trifluoro acetic        acid) in 5 min    -   System I: 60% acetonitrile, 40% water (0.1% trifluoro acetic        acid) to 100% acetonitrile in 5 min    -   System J: acetonitrile, 20% water (0.1% trifluoro acetic acid)        to 100% acetonitrile in 5 min    -   System K: 15% acetonitrile, 85% water (0.1% trifluoro acetic        acid) to 100% acetonitrile in 5 min

System L: acetonitrile, 30% water (0.1% trifluoro acetic acid) to 100%acetonitrile in 5 min TABLE F-3 LCMS parent peak and retention timevalues. Retention time Eluent Co. No. (minutes) LCMS [M + H] System 13.23 392 K 2 3.45 406 A 3 3.41 420 B 4 3.44 436 A 6 3.13 408 A 8 3.67443 D 9 3.26 464 K 15 3.42 422 B 16 3.78 404 A 17 3.33 380 K 18 3.86 426B 19 2.47 420 D 20 3.45 393 B 21 3.89 457 B 22 3.31 455 C 23 3.62 404 B24 3.58 450 A 25 2.23 478 F 26 3.58 378 A 27 2.90 394 B 28 3.31 380 C 293.58 479 D 30 3.32 379 A 31 2.48 464 D 32 3.34 462 C 33 3.07 393 C 353.15 469 K 36 3.39 418 B 37 3.43 436 A 38 3.50 464 A 39 4.00 518 A 403.58 418 A 41 3.32 505 D 42 2.74 564 D 43 3.50 464 A 44 3.11 592 D 453.77 492 A 46 3.25 678 E 47 3.30 478 B 48 3.35 508 A 49 2.77 408 B 503.54 495 A 51 3.17 395 A 52 3.55 405 K 53 2.82 466 B 54 2.74 438 B 553.33 538 A 56 4.11 407 B 57 3.54 432 B 58 3.64 478 A 59 3.38 406 A 603.37 420 A 61 2.97 606 L 62 3.92 520 A 63 3.40 478 A 64 3.99 548 A 653.56 448 A 66 3.84 507 K 67 3.57 407 B 68 3.90 532 A 69 3.51 432 A 703.33 538 A 71 3.00 438 A 72 3.30 522 A 73 2.97 422 A 74 3.79 550 A 753.36 450 A 76 3.83 578 A 77 3.43 478 A 79 3.05 422 A 80 3.00 436 K 812.80 396 K 83 2.78 436 B 84 3.33 442 A 85 3.47 459 B 86 3.00 422 A 873.00 436 A 88 3.17 420 A 89 2.60 407 A 90 3.11 465 B 95 4.23 421 B 963.71 450 A 97 3.64 4.92 B 98 3.57 492 B

1. A compound of formula (I),

a N-oxide, an addition salt, a quatemnary amine and a stereochemic allyisomeric form thereof, wherein X¹ and X² are each independently N or CHwith the exception that X¹ and X² can not be both N; Q¹ is CH₂, or N; Q²is CH₂, N, or O; n is an integer with value 0 or 1 and when n is 0 thena direct bond is intended; t is an integer with value 0 or 1 and when tis 0 then a direct bond is intended; -Y

Z- is —CH═N—, —C(═O)—NH—, —N═CH—, —C(═O)—CH₂—, or —NH—C(═O)—; ring Brepresents phenyl, cyclopentyl, cyclohexyl, norbornyl or

L is a direct bond, —(CH₂)_(r)—NR⁷—(CH₂)_(s)—, —(CR⁸ ₂)_(r—O—(CH)₂)_(s)—, —C(═O)—, —(CH₂)_(r)—O—C(═O)—, —(CH₂)_(r)—NR⁷—C(═O)—, —S(═O)₂—,—(CH₂)_(r)—NH—S(═O)₂—, or —C₁₋₆alkyl-; wherein each —(CH₂)_(r)— moietyis linked to ring A; each s is an integer with value 0 or 1 and when sis 0 then a direct bond is intended; each r is an integer with value 0,1, 2 or 3 and when r is 0 then a direct bond is intended; each R⁷ ishydrogen, C₁₋₆alkyl or C₁₋₄alkyloxycarbonyl; each R⁸ is independentlyhydrogen, hydroxy or C₁₋₆alkyl; or two R⁸ together can form a bivalentradical of formula —CH₂—CH₂—; R¹, R² and R⁵ are each independentlyhydrogen, hydroxy or C₁₋₆alkyl; R³ is hydrogen, hydroxy, C₁₋₆alkyl,hydroxyC₁₋₆alkyl, hydroxycyclopropylC₁₋₆alkyl,hydroxycyclopropylcarbonyl,hydroxyC₁₋₆alkylcarbonyl,hydroxyC₁₋₆alkyloxy, C₁₋₆alkyloxy,hydroxyC₁₋₆alkyloxyC₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkylcarbonyl,C₁₋₄alkyloxycarbonyl, C₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkyloxyC₁₋₆alkylcarbonyl, C₁₋₆alkyloxyC₁₋₆alkyloxy,C₁₋₆alkyloxyC₁₋₆alkyloxyC₁₋₆alkyl, pyridinyl, —NR⁹R¹⁰, or—S(═O)₂—NR⁹R¹⁰; wherein each R⁹ and R¹⁰ independently representhydrogen, C₁₋₆alkyl, C₁₋₄alkyloxycarbonyl, hydroxyC₁₋₆alkyl,hydroxycyclopropylC₁₋₆alkyl or C₁₋₆alkyloxyC₁₋₆alkyl; R⁴ is hydrogen orhalo; R⁶ is hydrogen, C₁₋₆alkyl or C₁₋₄alkyloxycarbonyl.
 2. A compoundas claimed in claim 1 wherein X¹ and X² are each CH.
 3. A compound asclaimed in claim 1 wherein n is 0; t is 0; L is a direct bond,—(CH₂)_(r)—NR⁷—(CH₂)_(s)—, —C(═O)—, —(CH₂)_(r)—NR⁷—C(═O)—, —S(═O)₂— or—C₁₋₆alkyl-; s is 1; r is 0 or 2; each R⁷ is hydrogen orC₁₋₄alkyloxycarbonyl; R¹, R² and R⁵ are each independently hydrogen; R³is hydrogen, hydroxy, C₁₋₆alkyl, hydroxyC₁₋₆alkyl,hydroxycyclopropylcarbonyl, hydroxyC₁₋₆alkylcarbonyl, C₁₋₆alkylcarbonyl,C₁₋₄alkyloxycarbonyl, C₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkyloxyC₁₋₆alkylcarbonyl, pyridinyl, —NR⁹R¹⁰, or —S(═O)₂—NR⁹R¹⁰;each R⁹ and R¹⁰ independently represent hydrogen, C₁₋₄alkyloxycarbonylor C₁₋₆alkyloxyC₁₋₆alkyl; and R⁶ is hydrogen or C₁₋₄alkyloxycarbonyl. 4.A compound as claimed in claim 1, wherein X¹ and X² are each CH; Q² isN, or O; n is 0; t is 0; -Y

Z- is —N═CH—; ring B represents cyclohexyl or norbornyl; L is a directbond, —(CH₂)_(r)—NR⁷—(CH₂)_(s)—, —C(═O)— or —C₁₋₆alkyl-; s is 1; r is 0or 2; each R⁷ is hydrogen; R¹, R² and R⁵ are each independentlyhydrogen; R³ is hydrogen, C₁₋₆alkyl or hydroxyC₁₋₆alkyl; R⁴ is hydrogen;and R⁶ is hydrogen.
 5. A compound as claimed in claim 1, wherein X¹ andX² are each CH; Q² is N, or O; n is 0; t is 0; -Y

Z- is —N═CH—; ring B represents cyclohexyl or norbornyl; L is a directbond, —NH—CH₂—, —C(═O)— or methyl; R¹, R² and R⁵ are each independentlyhydrogen; R⁴ is hydrogen; and R⁶ is hydrogen.
 6. A compound selectedfrom the group consisting of:

Compound No 1

Compound No 2

Compound No 15

Compound No 3

Compound No 60

Compound No 37

Compound No 4

Compound No 23


7. A pharmaceutical composition comprising pharmaceutically acceptablecarriers and as an active ingredient a therapeutically effective amountof a compound as claimed in claim
 1. 8. A process of preparing apharmaceutical composition as claimed in claim 7 wherein thepharmaceutically acceptable carriers and a compound as claimed in claim1 are intimately mixed. 9.-10. (canceled)
 11. A combination of ananti-cancer agent and a cell cycle kinase inhibitor as claimed in anyclaim
 1. 12. A process for preparing a compound as claimed in claim 1,said process comprising: a) reacting an intermediate of formula (II)with an intermediate of formula (III) in the presence of a suitablesolvent and optionally in the presence of a suitable base, resulting ina compound of formule (I),

b) reacting an intermediate of formula (X), wherein W¹ is a suitableleaving group, with an intermediate of formula (II) in the presence of asuitable solvent, optionally in the presence of a suitable base,resulting in the formation of a compound of formula (I),

c) reacting an intermediate of formula (XIV) with an intermediate offormula (XVI-a) in the presence of a suitable solvent optionally in thepresence of a suitable base with the formation of compounds of formula(I) wherein L is —(CH₂)_(r)—NH—C(═O)—, herein referred to as compoundsof formula (I-a),

d) reacting intermediates of formula (XIV) with intermediates of formula(XVI-b) in the presence of sodium triacetoxyborohydride or sodiumcyanoborohydride, in the presence of a suitable acid and in a suitablesolvent, resulting in compounds of formula (I) wherein L is—(CH₂)_(r)—NH—(CH₂)—, herein referred to as compounds of formula (I-b),and

e) cyclizing an intermediate of formula (XXXIII) in the presence of asalt or an acid in a suitable solvent, resulting in compounds of formula(I), wherein -Y

Z- is —CH═N—, herein refered to as compounds of formula (I-c).


13. A compound as claimed in claim 2 wherein n is 0; t is 0; L is adirect bond, —(CH₂)_(r)—NR⁷—(CH₂)_(s)—, —C(═O)—, —(CH₂)_(r)—NR⁷—C(═O)—,—S(═O)₂— or —C₁₋₆alkyl-; s is 1; r is 0 or 2; each R⁷ is hydrogen orC₁₋₄alkyloxycarbonyl; R¹, R² and R⁵ are each independently hydrogen; R³is hydrogen, hydroxy, C₁₋₆alkyl, hydroxyC₁₋₆alkyl,hydroxycyclopropylcarbonyl, hydroxyC₁₋₆alkylcarbonyl, C₁₋₆alkylcarbonyl,C₁₋₄alkyloxycarbonyl, C₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkyloxyC₁₋₆alkylcarbonyl, pyridinyl, —NR⁹R¹⁰, or —S(═O)₂—NR⁹R¹⁰;each R⁹ and R¹⁰ independently represent hydrogen, C₁₋₄alkyloxycarbonylor C₁₋₆alkyloxyC₁₋₆alkyl; and R⁶ is hydrogen or C₁₋₄alkyloxycarbonyl.14. A compound as claimed in claim 2 wherein X¹ and X² are each CH; Q²is N, or O; n is 0; t is ; -Y

Z- is —N═CH—; ring B represents cyclohexyl or norbornyl; L is a directbond, —(CH₂)_(r)—NR⁷—(CH₂)_(s)—, —C(═O)— or —C₁₋₆alkyl-; s is 1; r is 0or 2; each R⁷ is hydrogen; R¹,R² and R⁵ are each independently hydrogen;R³ is hydrogen, C₁₋₆alkyl or hydroxyC₁₋₆alkyl; R⁴ is hydrogen; and R⁶ ishydrogen.
 15. A compound as claimed in claim 3 wherein X¹ and X² areeach CH; Q² is N, or O; n is 0; t is ; -Y

Z- is —N═CH—; ring B represents cyclohexyl or norbornyl; L is a directbond, —(CH₂)_(r)—NR⁷—(CH₂)_(s)—, —C(═O)— or —C₁₋₆alkyl-; s is 1; r is 0or 2; each R⁷ is hydrogen; R¹, R² and R⁵ are each independentlyhydrogen; R³ is hydrogen, C₁₋₆alkyl or hydroxyC₁₋₆alkyl; R⁴ is hydrogen;and R⁶ is hydrogen.
 16. A compound as claimed in claim 2 wherein X¹ andX² are each CH; Q² is N, or O; n is 0; t is 0; -Y

Z- is —N═CH—; ring B represents cyclohexyl or norbornyl; L is a directbond, —NH—CH₂—, —C(═O)— or methyl; R¹, R² and R⁵ are each independentlyhydrogen; R⁴ is hydrogen; and R⁶ is hydrogen.
 17. A compound as claimedin claim 3 wherein X¹ and X² are each CH; Q² is N, or O; n is 0; t is 0;-Y

Z- is —N═CH—; ring B represents cyclohexyl or norbornyl; L is a directbond, —NH—CH₂—, —C(═O)— or methyl; R¹, R² and R⁵ are each independentlyhydrogen; R⁴ is hydrogen; and R⁶ is hydrogen.
 18. A compound as claimedin claim 4 wherein X¹ and X² are each CH; Q² is N, or O; n is 0; t is 0;-Y

Z- is —N═CH—; ring B represents cyclohexyl or norbomyl; L is a directbond, —NH—CH₂—, —C(═O)— or methyl; R¹, R² and R⁵ are each independentlyhydrogen; R⁴ is hydrogen; and R⁶ is hydrogen.
 19. A pharmaceuticalcomposition comprising pharmaceutically acceptable carriers and as anactive ingredient a therapeutically effective amount of a compound asclaimed in claim
 6. 20. A process of preparing a pharmaceuticalcomposition as claimed in claim 19 wherein the pharmaceuticallyacceptable carriers and a compound as claimed in claim 6 are intimatelymixed.
 21. A combination of an anti-cancer agent and a cell cycle kinaseinhibitor as claimed in any claim
 6. 22. A method of treating in asubject proliferative diseases or differentiative disorders comprisingadministering to the subject a therapeutically effective amount of apharmaceutical composition comprising a compound of claim 1 and apharmaceutically acceptable carrier.
 23. A method of treating in asubject proliferative diseases or differentiative disorders comprisingadministering to the subject a therapeutically effective amount of apharmaceutical composition comprising a compound of claim 6 and apharmaceutically acceptable carrier.